Retrovirus from the HIV group and its use

ABSTRACT

A novel immunodeficiency virus is disclosed which has the designation MVP-5180/91 and which has been deposited with the European Collection of Animal Cell Cultures (ECACC) under No. V 920 52 318. The characteristics antigens which can be obtained from it and which can be employed for detecting antibodies against retroviruses which are associated with immunodeficiency diseases are also disclosed, as are the DNA and amino acid sequences of the virus.

[0001] This application is a continuation of U.S. application Ser. No.09/109,916, filed Jul. 2, 1998, now allowed, which is a divisionalapplication of U.S. application Ser. No. 08/468,059, filed Jun. 6, 1995,now U.S. Pat. No. 5,840,480, issued Nov. 24, 1998; which is a divisionalapplication of U.S. application Ser. No. 08/132,653, filed Oct. 5, 1993,now abandoned; the disclosures of all of which are incorporated hereinby reference.

[0002] The present invention relates to a novel retrovirus from the HIVgroup, as well as to variants or parts thereof which contain theessential properties of the virus. A process is described for culturingthe retrovirus. The invention furthermore relates to the isolation ofthis retrovirus and to use of the virus, its parts or extracts formedicinal purposes, for diagnostics and in the preparation of vaccines.

[0003] Retroviruses which belong to the so-called HIV group lead inhumans who are infected by them to disease manifestations which aresummarized under the collective term immunodeficiency or AIDS (acquiredimmune deficiency syndrome).

[0004] Epidemiological studies verify that the human immunodeficiencyvirus (HIV) represents the etiological agent in the vast majority ofAIDS (acquired immune deficiency syndrome) cases. A retrovirus which wasisolated from a patient and characterized in 1983 received thedesignation HIV-1 (Barré-Sinoussi, F. et al., Science 220, 868-871[1983]). A variant of HIV-1 is described in WO 86/02383.

[0005] A second group of human immunodeficiency viruses was identifiedin 1985 in West Africa (Clavel, F. et al., Science 233, 343-346 [1986])and designated human immunodeficiency virus type 2 (HIV-2) (EP-A-0 239425). While HIV-2 retroviruses clearly differ from HIV-1, they doexhibit affinity with simian immunodeficiency viruses (SIV-2). LikeHIV-1, HIV-2 also leads to AIDS symptomatology.

[0006] A further variant of an immunodeficiency retrovirus is describedin EP-A-0 345 375 and designated there as HIV-3 retrovirus (ANT 70).

[0007] The isolation of a further, variant, immunodeficiency virus isalso described in Lancet Vol. 340, September 1992, pp. 681-682.

[0008] It is characteristic of human immunodeficiency viruses that theyexhibit a high degree of variability, which significantly complicatesthe comparability of the different isolates. For example, when diverseHIV-1 isolates are compared, high degrees of variability are found insome regions of the genome while other regions are comparatively wellconserved (Benn, S. et al., Science 230, 949-951 (1985]). It was alsopossible to observe an appreciably greater degree of polymorphism in thecase of HIV-2 (Clavel, F. et al., Nature 324, 691-695 [1986]). Thegreatest degree of genetic stability is possessed by regions in the gagand pol genes which encode proteins which are essential for structuraland enzymic purposes; some regions in the env gene, and the genes (vif,vpr, tat, rev and nef) encoding regulatory proteins, exhibit a highdegree of variability. In addition to this, it was possible todemonstrate that antisera against HIV-1 also crossreact with gag and polgene products from HIV-2 even though there was only a small degree ofsequence homology. Little hybridization of significance likewise tookplace between these two viruses unless conditions of very low stringencywere used (Clavel, F. et al., Nature 324, 691-695 [1986]).

[0009] Owing to the wide distribution of retroviruses from the HIV groupand to the fact that a period of a few to many years (2-20) existsbetween the time of infection and the time at which unambiguous symptomsof pathological changes are recognizable, it is of great importance fromthe epidemiological point of view to determine infection withretroviruses of the HIV group at as early a stage as possible and, aboveall, in a reliable manner. This is not only of importance whendiagnosing patients who exhibit signs of immunodeficiency, but also whenmonitoring blood donors. It has emerged that, when retroviruses of theHIV-1 or HIV-2 type, or components thereof, are used in detectionsystems, antibodies can either not be detected or only detected weaklyin many sera even though signs of immunodeficiency are present in thepatients from which the sera are derived. In certain cases, suchdetection is possible using the retrovirus from the HIV group accordingto the invention.

[0010] This patent describes the isolation and characterization of anovel human immunodeficiency virus, designated below as MVP-5180/91 (SEQID NO:56), which was isolated from the peripheral lymphocytes of afemale patient from the Cameroons who was 34 years old in 1991 and whoexhibited signs of immunodeficiency. From the point of view ofgeography, this retrovirus originates from a region in Africa which islocated between West Africa, where there is endemic infection with HIV-2and HIV-1 viruses, and Eastern Central Africa, where it is almostexclusively HIV-1 which is disseminated. Consequently, the presentinvention relates to a novel retrovirus, designated MVP-5180/91 (SEQ IDNO:56), of the HIV group and its variants, to DNA sequences, amino acidsequences and constituent sequences derived therefrom, and to test kitscontaining the latter. The retrovirus MVP-5180/91 (SEQ ID NO:56) hasbeen deposited with the European Collection of Animal Cell Cultures(ECACC) PHLS Centre for Applied Microbiology & Research, Porton Down,Salisbury Wilts. SP4 OJG, United Kingdom, on Sep.23, 1992 under ECACCAccession No. V 920 92 318 in accordance with the stipulations of theBudapest Treaty. The ECACC is located at the PHLS Centre for AppliedMicrobiology & Research, Porton Down, Salisbury, Wilts, SP4 OJG, U.K.The deposit was made on Sep. 23, 1992, and was assigned Accession No. V920 92 318. The date of notification of acceptance of the culture wasJan. 21, 1993.

[0011] As do HIV-1 and HIV-2, MVP-5180/91 (SEQ ID NO:56) according tothe invention grows in the following cell lines: HUT 78, Jurkat cells,C8166 cells and MT-2 cells. The isolation and propagation of viruses isdescribed in detail in the book “Viral Quantitation in HIV Infection,Editor Jean-Marie Andrieu, John Libbey Eurotext, 1991”. The proceduralmethods described in that publication are by reference made a subject ofthe disclosure of the present application.

[0012] In addition to this, the virus according to the inventionpossesses a reverse transcriptase which is magnesium-dependent but notmanganese-dependent. This represents a further property possessed incommon with the HIV-1 and HIV-2 viruses.

[0013] In order to provide a better understanding of the differencesbetween the MVP-5180/91 (SEQ ID NO:56) virus according to the inventionand the HIV-1 and HIV-2 retroviruses, the construction of theretroviruses which cause immunodeficiency will first of all be explainedin brief. Within the virus, the RNA is located in a conical core whichis assembled from protein subunits which carry the designation p 24 (pfor protein). This inner core is surrounded by a protein coat, which isconstructed from protein p 17 (outer core), and by a glycoprotein coatwhich, in addition to lipids, which originate from the host cell,contains the transmembrane protein gp 41 and the coat protein 120 (gp120). This gp 120 can then bind to the CD-4 receptors of the host cells.

BRIEF DESCRIPTION OF THE DRAWINGS

[0014]FIG. 1 depicts the arrangement of the genome of retroviruses ofthe HIV type.

[0015]FIG. 2 is a graph depicting the binding affinity for themonoclonal antibody p24 in relation to the content of reversetranscriptase for the retroviruses HIV-1, HIV-2, and MVP-5180/91.

[0016]FIG. 3 depicts a western blot of MVP-5180/91 and HIV-1, isolatedfrom German patients.

[0017]FIG. 4 depicts the almost complete nucleic acid sequence of theretrovirus MVP-5180/91.

[0018]FIG. 5 depicts the strategy for PCR amplification, cloning, andsequencing of MVP-5180/91.

[0019]FIG. 6 depicts a comparison of the sequence in FIG. 4 and thesequence obtained using the PCR amplification techniques depicted inFIG. 5.

[0020]FIG. 7 depicts a comparison of the amino acid sequences of the GAGprotein determined from the sequence of FIG. 4 with the GAG proteinsequence obtained using the PCR amplification techniques depicted inFIG. 5.

[0021]FIG. 8 depicts the immunological specificities of the V3 loop ofHIV-1, HIV-2, and MVP-5180/91.

DETAILED DESCRIPTION OF THE INVENTION

[0022] As far as is known, the RNA of HIV viruses—portrayed in asimplified manner—possesses the following gene regions: so-called longterminal repeats (LTR) at each end, together with the following generegions: gag, pol, env and nef. The gag gene encodes, inter alia, thecore proteins, p 24 and p 17, the pol gene encodes, inter alia, thereverse transcriptase, the RNAse H and the integrase, while the env geneencodes the gp 41 and gp 120 glycoproteins of the virus coat. The nefgene encodes a protein having a regulatory function. The arrangement ofthe genome of retroviruses of the HIV type is shown diagrammatically inFIG. 1.

[0023] The HIV-1 and HIV-2 retroviruses can be distinguished, interalia, by testing viral antigen using a monoclonal antibody which iscommercially available from Abbott (HIVAG-1 monoclonal) in the form of atest kit and is directed against (HIV-1) p 24. It is known that thecontent of reverse transcriptase is roughly the same in the HIV-1 andHIV-2 virus types. If, therefore, the extinction (E 490 nm.) obtained indilutions of the disrupted viruses by means of the antigen-antibodyreaction is plotted against the activity of reverse transcriptase, aseries of graphs is obtained corresponding roughly to that in FIG. 2. Inthis context, it is observed that, in the case of HIV-1, the monoclonalantibody employed has a very high binding affinity for p 24 in relationto the content of reverse transcriptase. By contrast, the monoclonalantibody employed has only a very low binding affinity for p 24 in thecase of HIV-2, once again in relation to the content of reversetranscriptase. If these measurements are carried out on MVP-5180/91 (SEQID NO:56), the curve is then located almost precisely in the centrebetween the curves for HIV-1 and HIV-2, i.e. the binding affinity of themonoclonal antibody for MVP-5180/91 p 24 is reduced as compared with thecase of HIV-1. FIG. 2 shows this relationship diagrammatically, with RTdenoting reverse transcriptase, and the protein p 24, against which isdirected the monoclonal antibody which is present in the test kit whichcan be purchased from Abbott, being employed as the antigen (Ag).

[0024] The so-called PCR (polymerase chain reaction) system has provedto have a multiplicity of uses in genetic manipulation, and thecomponents which are required for implementing the process can bepurchased. Using this process, it is possible to amplify DNA sequencesif regions of the sequence to be amplified are known. Short,complementary DNA fragments (oligonucleotides=primers) have then to besynthesized which anneal to a short region of the nucleic acid sequenceto be amplified. For carrying out the test, HIV nucleic acids areintroduced together with the primers into a reaction mixture whichadditionally contains a polymerase and nucleotide triphosphates. Thepolymerization (DNA synthesis) is carried out for a given time and thenucleic acid strands are then separated by heating. After cooling, thepolymerization then proceeds once more. If, therefore, the retrovirusaccording to the invention is an HIV-1 or HIV-2 virus, it should bepossible to amplify the nucleic acid using primers which are conservedwithin the known sequences of the HIV-1 and HIV-2 viruses. Some primersof this type have previously been described (Lauré, F. et al., Lancetii, (1988) 538-541 for pol 3 and pol 4, and Ou C.Y. et al., Science 239(1988) 295-297 for sk 38/39 and sk 68/69).

[0025] It was discovered that use of particular primer pairs having thefollowing sequence: gaga (SEQ ID CTACT AGTAC CCTTC AGG NO:1): gagb (SEQID CGGTC TACAT AGTCT CTAAA G NO:2): sk38 (SEQ ID CCACC TATCC CAGTA GGAGAA NO:3): sk39 (SEQ ID CCTTT GGTCC TTGTC TTATG TCCAG AATGC NO:4): or po13(SEQ ID TGGGA AGTTC AATTA GGAAT ACCAC NO:5): po14 (SEQ ID CCTAC ATAGAAATCA TCCAT GTATT G NO:6): pol3n (SEQ ID TGGAT GTGGG TGATG CATA NO:7):pol4n (SEQ ID AGCAC ATTGT ACTGA TATCT A NO:8): and SK145 (SEQ ID AGTGGGGGGA CATCA AGCAG CC NO:9): SK150 (SEQ ID TGCTA TGTCA CTTCC CCTTG GTNO:l0): 145-P (SEQ ID CCATG CAAAT GTTAA AAGAG AC NO:11): 150-P (SEQ IDGGCCT GGTGC AATAG GCCC NO:12): or a combination of pol 3 and pol 4 withUNI-1 (SEQ ID GTGCT TCCAC AGGGA TGGAA NO:13): UNI-2 (SEQ ID ATCAT CCATGTATTG ATA NO:14):

[0026] (Donehower L. A. et al. (1990) J. Virol. Methods 28, 33-46) andemploying PCR with nested primers, led to weak amplifications of theMVP-5180/91 DNA (SEQ ID NO:56).

[0027] No amplification, or only weak amplification as compared withHIV-1, possibly attributable to impurities, was obtained with thefollowing primer sequences: tat 1: (SEQ ID NO:15) AATGG AGCCA GTAGATCCTA tat 2: (SEQ ID NO:16) TGTCT CCGCT TCTTC CTGCC tat 1P: (SEQ IDNO:17): GAGCC CTGGA AGCAT CCAGG tat 2P: (SEQ ID NO:18): GGAGA TGCCTAAGGC TTTTG enva: (SEQ ID NO:19): TGTTC CTTGG GTTCT TG envb: (SEQ IDNO:20): GAGTT TTCCA GAGCA ACCCC sk68: (SEQ ID NO:21): AGCAG CAGGA AGCACTATGG sk69: (SEQ ID NO:22): GCCCC AGACT GTGAG TTGCA ACAG 5v3e: (SEQ IDNO:23): GCACA GTACA ATGTA CACAT GG 3v3e: (SEQ ID NO:24): CAGTA GAAAAATTCC CCTCC AC 5v3degi: (SEQ ID NO:25): TCAGG ATCCA TGGGC AGTCT AGCAGAAGAA G 3v3degi: (SEQ ID NO:26): ATGCT CGAGA ACTGC AGCAT CGATT CTGGGTCCCC TCCTG AG 3v3longdegi: (SEQ ID NO:27): CGAGA ACTGC AGCAT CGATGCTGCT CCCAA GAACC CAAGG 3v3longext: (SEQ ID NO:28): GGAGC TGCTT GATGCCCCAG A gagdi: (SEQ ID NO:29): TGATG ACAGC ATGTC AGGGA GT pol e: (SEQ IDNO:30): GCTGA CATTT ATCAC AGCTG GCTAC tat 1 (SEQ ID NO:15): AATGG AGCCAGTAGA TCCTA

[0028] Amplifications which were weak as compared with those for HIV-1,but nevertheless of the same intensity as those for the HIV-2 isolate(MVP-11971/87) employed, were obtained with gag c: (SEQ ID NO: TATCACCTAG AACTT TAAAT GCATG GG 31): gag d: (SEQ ID NO: AGTCC CTGAC ATGCTGTCAT CA 32): env c: (SEQ ID NO: GTGGA GGGGA ATTTT TCTAC TG 33): env d:(SEQ ID NO: CCTGC TGCTC CCAAG AACCC AAGG. 34):

[0029] The so-called Western blot (immunoblot) is a common method fordetecting HIV antibodies. In this method, the viral proteins arefractionated by gel electrophoresis and then transferred to a membrane.The membranes provided with the transferred proteins are then broughtinto contact with sera from the patients to be investigated. Ifantibodies against the viral proteins are present, these antibodies willbind to the proteins. After the membranes have been washed, onlyantibodies which are specific for the viral proteins will remain. Theantibodies are then rendered visible using antiantibodies which, as arule, are coupled to an enzyme which catalyzes a color reaction. In thisway, the bands of the viral proteins can be rendered visible.

[0030] The virus MVP-5180/91 (SEQ ID NO:56) according to the inventionexhibits two significant and important differences from the HIV-1 andHIV-2 viruses in a Western blot. HIV-1 regularly shows a strong band,which is attributable to protein p 24, and a very weak band, which isoften scarcely visible and which is attributable to protein p 23. HIV-2exhibits a strong band, which is attributable to protein p 25, andsometimes a weak band, which is attributable to protein p 23. Incontrast to this, the MVP-5180/91 (SEQ ID NO:56) virus according to theinvention exhibits two bands of approximately equal strength,corresponding to proteins p 24 and p 25.

[0031] A further significant difference exists in the bands which areattributable to reverse transcriptase. HIV-1 shows one band (p 53) whichcorresponds to reverse transcriptase and one band (p 66) whichcorresponds to reverse transcriptase bound to RNAse H. In the case ofHIV-2, the reverse transcriptase corresponds to protein p 55 and, if itis bound to RNAse H, to protein p 68. By contrast, MPV-5180/91 (SEQ IDNO:56) according to the invention exhibits one band at protein p 48,which corresponds to reverse transcriptase, and one band, at protein p60, which corresponds to reverse transcriptase bound to RNAse H. It canbe deduced from these results that the reverse transcriptase ofMVP-5180/91 (SEQ ID NO:56) has a molecular weight which is roughlybetween 3 and 7 kilodaltons less than that of the reverse transcriptasesof HIV-1 and HIV-2. The reverse transcriptase of MVP-5180 consequentlyhas a molecular weight which is roughly between 4,500 daltons and 5,500daltons less than that of the reverse transcriptase of HIV-1 or HIV-2.

[0032] It was discovered that anti-env antibodies could only be detectedweakly in the sera of German patients exhibiting signs ofimmunodeficiency when the MVP-5180/91 (SEQ ID NO:56) virus according tothe invention was used, whereas the sera reacted strongly if an HIV-1virus was used instead of the virus according to the invention. Thisstronger detection reaction was located in the gp 41 protein, inparticular. In the experiments, serum panels were compared which on theone hand derived from German patients and on the other from Africanpatients showing signs of immune deficiency.

[0033] The abovementioned characteristics are indicative of those virusvariants which correspond to MVP-5180/91 (SEQ ID NO:56) according to theinvention. Therefore, the virus according to the invention, or variantsthereof, can be obtained by isolating immunodeficiency viruses fromheparinized donor blood derived from persons who exhibit signs of immunedeficiency and who preferably originate from Africa.

[0034] Since the virus possessing the abovementioned properties has beenisolated, the cloning of a cDNA can be carried out in the followingmanner: the virus is precipitated from an appropriately large quantityof culture (about 1 l) and then taken up in phosphate-buffered sodiumchloride solution. It is then pelleted through a (20% strength) sucrosecushion. The virus pellet can be suspended in 6 M guanidinium chloridein 20 mM dithiothreitol and 0.5% Nonidet P 40. CsCl is added to bringits concentration to 2 molar and the solution containing the disruptedvirus is transferred to a cesium chloride cushion. The viral RNA is thenpelleted by centrifugation, and subsequently dissolved, extracted withphenol and precipitated with ethanol and lithium chloride. Synthesis ofthe first cDNA strand is carried out on the viral RNA, or parts thereof,using an oligo(dT) primer. The synthesis can be carried out using acommercially available kit and adding reverse transcriptase. Tosynthesize the second strand, the RNA strand of the RNA/DNA hybrid isdigested with RNase H, and the second strand is then synthesized usingE. coli DNA polymerase I. Blunt ends can then be produced using T4 DNApolymerase and these ends can be joined to suitable linkers forrestriction cleavage sites. Following restriction digestion with theappropriate restriction endonuclease, the cDNA fragment is isolated froman agarose gel and ligated to a vector which has previously been cut inan appropriate manner. The vector containing the cDNA insert can then beused for transforming competent E. coli cells. The colonies which areobtained are then transferred to membranes, lysed and denatured, andthen finally detected by hybridization with nucleic acid labeled withdigoxigenin or biotin. Once the corresponding cDNA has been prepared bygenetic manipulation, it is possible to isolate the desired DNAfragments originating from the retrovirus. By incorporating thesefragments into suitable expression vectors, the desired protein orprotein fragment can then be expressed and employed for the diagnostictests.

[0035] As an alternative to the stated method, the immunodeficiencyvirus can be cloned with the aid of PCR technology, it being possible touse the abovementioned primers.

[0036] The similarity between different virus isolates can be expressedby the degree of homology between the nucleic acid or protein sequences.50% homology means, for example, that 50 out of 100 nucleotides or aminoacid positions in the sequences correspond to each other. The homologyof proteins is determined by sequence analysis. Homologous DNA sequencescan also be identified by the hybridization technique.

[0037] In accordance with the invention, a part of the coat protein wasinitially sequenced and it was ascertained that this sequence possessedonly relatively slight homology to the corresponding sequences fromviruses of the HIV type. On the basis of a comparison with HIVsequences, which was carried out using data banks, it was established,in relation to the gp 41 region in particular, that the homology was atmost 66% (nucleotide sequence).

[0038] In addition to this, the region was sequenced which encodes gp41. This sequence is presented in Tables 1 and 3. Table 1 includes DNASEQ ID NO:37, DNA SEQ ID NO:38, and amino acid SEQ ID NO:39. Table 3includes DNA SEQ ID NO:44, DNA SEQ ID NO:45, and amino acid SEQ IDNO:46.

[0039] The present invention therefore relates to those viruses whichpossess an homology of more than 66%, preferably 75% and particularlypreferably 85%, to the HIV virus, MVP-5180/91 (SEQ ID NO:56), accordingto the invention, based on the nucleotide sequence in Table 1 (SEQ IDNO:37; SEQ ID NO:38) and/or in Table 3 (SEQ ID NO:44; SEQ ID NO:45).

[0040] Furthermore, the present invention relates to those viruses whichpossess an homology of more than 66%, preferably 75% and particularlypreferably 85%, to partial sequences of the nucleotide sequencepresented in Table 3 (SEQ ID NO:44; SEQ ID NO:45), which sequences areat least 50, preferably 100, nucleotides long. This corresponds to alength of the peptides of at least 16, and preferably of at least 33,amino acids.

[0041] The sequence of the virus according to the invention differs fromthat of previously known viruses. The present invention thereforerelates to those viruses, and corresponding DNA and amino acidsequences, which correspond to a large extent to the sequence of thevirus according to the invention, the degree of deviation beingestablished by the degree of homology. An homology of, for example, morethan 85% denotes, therefore, that those sequences are included whichhave in at least 85 of 100 nucleotides or amino acids the samenucleotides or amino acids, respectively, while the remainder can bedifferent. When establishing homology, the two sequences are compared insuch a way that the greatest possible number of nucleotides or aminoacids corresponding to each other are placed in congruence.

[0042] The (almost) complete sequence, given as the DNA sequence of thevirus according to the invention, is reproduced in FIG. 4 and includedas DNA SEQ ID NO:56. In this context, the present invention relates toviruses which possess the sequence according to FIG. 4 (SEQ ID NO:56),and variants thereof which possess a high degree of homology with thesequence of FIG. 4 (SEQ ID NO:56), as well as proteins, polypeptides andoligopeptides derived therefrom which can be used diagnostically or canbe employed as vaccines.

[0043] Using the isolated sequence as a basis, immunodominant epitopes(peptides) can be designed and synthesized. Since the nucleic acidsequence of the virus is known, the person skilled in the art can derivethe amino acid sequence from this known sequence. A constituent regionof the amino acid sequence is given in Table 3 (SEQ ID NO:46). Thepresent invention also relates, therefore, to antigens, i.e. proteins,oligopeptides or polypeptides, which can be prepared with the aid of theinformation disclosed in FIG. 4 (SEQ ID NO:56) and Table 3 (SEQ IDNO:44; SEQ ID NO:45, and SEQ ID NO:46). These antigens, proteins,polypeptides and oligopeptides possess amino acid sequences which caneither be derived from FIG. 4 (SEQ ID NO:56) or are given in Table 3(SEQ ID NO:46). The antigens or peptides can possess relatively shortconstituent sequences of an amino acid sequence which is reproduced inTable 3 (SEQ ID NO:46) or which can be derived from FIG. 4 (SEQ IDNO:56). This amino acid sequence is at least 6, preferably at least 10and particularly preferably at least 15, amino acids in length. Thesepeptides can be prepared not only with the aid of recombinant technologybut also using synthetic methods. A suitable preparation route issolid-phase synthesis of the Merrifield type. Further description ofthis technique, and of other processes known to the state of the art,can be found in the literature, e.g. M. Bodansky, et al., PeptideSynthesis, John Wiley & Sons, 2nd Edition 1976.

[0044] In the diagnostic tests, a serum sample from the person to beinvestigated is brought into contact with the protein chains of one ormore proteins or glycoproteins (which can be expressed in eukaryoticcell lines), or parts thereof, which originate from MVP-5180/91 (SEQ IDNO:56). Test processes which are preferred include immunofluorescence orimmunoenzymatic test processes (e.g. ELISA or immunoblot).

[0045] In the immunoenzymatic tests (ELISA), antigen originating fromMVP-5180/91 (SEQ ID NO:56) or a variant thereof, for example, can bebound to the walls of microtiter plates. The dosage used in this contextdepends to an important degree on the test system and the treatment ofthe microtiter plates. Serum or dilutions of serum deriving from theperson to be investigated are then added to the wells of the microtiterplates. After a predetermined incubation time, the plate is washed andspecific immunocomplexes are detected by antibodies which bindspecifically to human immunoglobulins and which had previously beenlinked to an enzyme, for example horseradish peroxidase, alkalinephosphatase, etc., or to enzyme-labeled antigen. These enzymes are ableto convert a colorless substrate into a strongly colored product, andthe presence of specific anti-HIV antibodies can be gathered from thestrength of the coloration. A further option for using the virusaccording to the invention in test systems is its use in Western blots.

[0046] Even if the preparation of vaccines against immunodeficiencydiseases is proving to be extremely difficult, this virus, too, or partsthereof, i.e. immunodominant epitopes and inducers of cellular immunity,or antigens prepared by genetic manipulation, can still be used fordeveloping and preparing vaccines.

EXAMPLE 1

[0047] The immunodeficiency virus according to the invention,MVP-5180/91 (SEQ ID NO:56), was isolated from the blood of a femalepatient exhibiting signs of immune deficiency. To do this, peripheralmononuclear cells (peripheral blood lymphocytes, PBL) and peripherallymphocytes from the blood (PBL) of a donor who was not infected withHIV were stimulated with phytohemagglutinin and maintained in culture.For this purpose, use was made of the customary medium RPMI 1640containing 10% fetal calf serum. The culture conditions are described inLanday A. et al., J. Inf. Dis., 161 (1990) pp. 706-710. The formation ofgiant cells was then observed under the microscope. The production ofHIV viruses was ascertained by determining the p 24 antigen using thetest which can be purchased from Abbott. An additional test fordetermining the growth of the viruses consisted of the test usingparticle-bound reverse transcriptase (Eberle J., Seibl R., J. Virol.Methods 40, 1992, pp. 347-356). The growth of the viruses was thereforedetermined once or twice a week on the basis of the enzymatic activitiesin the culture supernatant, in order to monitor virus production. Newdonor lymphocytes were added once a week.

[0048] Once it was possible to observe HIV virus multiplication, freshperipheral lymphocytes from the blood (PBL) of healthy donors, who werenot infected with HIV, were infected with supernatant from the firstculture. This step was repeated and the supernatant was then used toinfect H 9 and HUT 78 cells. In this way, it was possible to achievepermanent production of the immunodeficiency virus. The virus wasdeposited with the ECACC under No. V 920 92 318.

EXAMPLE 2

[0049] So-called Western blot or immunoblot is currently a standardmethod for detecting HIV infections. Various sera were examined inaccordance with the procedure described by Gürtler et al. in J. Virol.Meth. 15 (1987) pp. 11-23. In doing this, sera from German patients werecompared with sera which had been obtained from African patients. Thefollowing results were obtained: Virus type German sera African seraHIV-1, virus strong reaction strong reaction isolated from using gp 41German patients MVP-5180/91 (SEQ ID no reaction to strong reactionNO:56) weak reaction using gp 41

[0050] The results presented above demonstrate that a virus of the HIV-1type isolated from German patients may possibly, if used for detectingHIV infections, fail to provide unambiguous results if the patient wasinfected with a virus corresponding to MVP-5180/91 (SEQ ID NO:56)according to the invention. It is assumed here that those viruses can bedetected using the virus according to the invention which possess atleast about 85% homology, based on the total genome, with the virusaccording to the invention.

EXAMPLE 3

[0051] Further Western blots were carried out in accordance with theprocedure indicated in Example 2. The results are presented in theenclosed FIG. 3. In this test, the viral protein of the immunodeficiencyvirus MVP-5180/91 (SEQ ID NO:56) according to the invention, in the onecase, and the viral protein of an HIV-1 type virus (MVP-899), in theother, was fractionated by gel electrophoresis and then transferred tocellulose filters. These filter strips were incubated with the sera fromdifferent patients and the specific antibodies were then renderedvisible by a color reaction. The left half of the figure with theheading MVP-5180 shows the immunodeficiency virus according to theinvention. The right half of the figure shows a virus (MVP-899), whichis an HIV-1 virus, isolated from a German donor.

[0052] In FIG. 3, the same sera (from German patients) were in each casereacted with two respective filter strips, the numbers 8 and 26; 9 and27; 10 and 28; 11 and 29; 12 and 30; 13 and 31; 14 and 32; 15 and 33,and 16 and 34 indicating the same sera. Sera from African patients wereemployed in the Western blots having the numbers 17 and 18. The numberson the right hand margins indicate the approximate molecular weights inthousands (KD).

[0053]FIG. 3 shows clearly that sera from German patients only reactvery weakly with the immunodeficiency virus according to the inventionin a Western blot using gp 41. By contrast, sera from African patientsreact very strongly with the immunodeficiency virus according to theinvention. FIG. 3 makes it clear, therefore, that when theimmunodeficiency virus according to the invention is used thoseimmunodeficiency infections can be detected which only yieldquestionable, i.e. not unambiguously positive, results when an HIV-1 orHIV-2 virus is used. This option for detection can be of far-reachingdiagnostic importance since, in those cases in which only questionableresults are obtained in a Western blot, it cannot be established withunambiguous certainty whether an infection with an immunodeficiencyvirus is present. However, if the immunodeficiency virus according tothe invention can be used to assign such questionable results to aninfection with a virus of the type according to the invention, this thenrepresents a substantial diagnostic advance.

EXAMPLE 4

[0054] DNA isolation, amplification and structural characterization ofsections of the genome of the HIV isolate MVP-5180/91 (SEQ ID NO:56).

[0055] Genomic DNA from HUT 78 cells infected with MVP-5180/91 (SEQ IDNO:56) was isolated by standard methods.

[0056] In order to characterize regions of the genome of the isolateMVP-5180/91 (SEQ ID NO:56), PCR (polymerase chain reaction) experimentswere carried out using a primer pair from the region of the coat proteingp 41. The PCR experiments were carried out in accordance with themethod of Saiki et al. (Saiki et al., Science 239: 487-491, 1988) usingthe following modifications: for the amplification of regions ofHIV-specific DNA, 5 μl of genomic DNA from HUT 78 cells infected withMVP-5180/91 (SEQ ID NO:56) were pipetted into a 100 μl reaction mixture(0.25 mM dNTP, in each case 1 μm primer 1 and primer 2, 10 mM Tris HCl,pH 8.3, 50 mM KCl, 1.5 MgCl₂, 0.001% gelatin, 2.5 units of Taqpolymerase (Perkin Elmer)), and amplification was then carried out inaccordance with the following temperature program: 1. initialdenaturation: 3′ 95° C., 2. amplification: 90″ 94° C., 60″ 56° C., 90″,72° C. (30 cycles).

[0057] The primers used for the PCR and for nucleotide sequencing weresynthesized on a Biosearch 8750 oligonucleotide synthesizer.

[0058] Primer 1 (SEQ ID NO:35): AGC AGC AGG AAG CAC TAT GG (coordinatesfrom HIV-1 isolate HXB2: bases 7795-7814, corresponds to primer sk 68)(SEQ ID NO:21)

[0059] Primer 2 (SEQ ID NO:36): GAG TTT TCC AGA GCA ACC CC (coordinatesfrom HIV-1 isolate HXB2: bases 8003-8022, corresponds to primer env b(SEQ ID NO:20).

[0060] The amplified DNA was fractionated on a 3% “Nusieve” agarose gel(from Biozyme) and the amplified fragment was then cut out and an equalvolume of buffer (1*TBE (0.09 M Tris borate, 0.002 M EDTA, pH 8.0) wasadded to it. After incubating the DNA/agarose mixture at 70° C. for 10minutes, and subsequently extracting with phenol, the DNA wasprecipitated from the aqueous phase by adding {fraction (1/10)} vol of 3M NaAc, pH 5.5, and 2 vol of ethanol and storing at −20° C. for 15′, andthen subsequently pelleted in a centrifuge (Eppendorf) (13,000 rpm, 10′,4° C.). The pelleted DNA was dried and taken up in water, and then,after photometric determination of the DNA concentration at 260 nm in aspectrophotometer (Beckman), sequenced by the Sanger method (F. Sanger,Proc. Natl. Acad,. Sci., 74: 5463, 1977). Instead of sequencing withKlenow DNA polymerase, the sequencing reaction was carried out using akit from Applied Biosystems (“Taq dye deoxy terminator cyclesequencing”, order No.: 401150). Primer 1 (SEQ ID NO:35) or primer 2(SEQ ID NO:36) (in each case 1 μM) was employed as primers in separatesequencing reactions. The sequencing reaction was analysed on a 373A DNAsequencing apparatus (Applied Biosystems) in accordance with theinstructions of the apparatus manufacturer.

[0061] The nucleotide sequence of the amplified DNA region, and theamino acid sequence deduced from it, are presented in Table 1. Table 1includes the DNA sequences SEQ ID NO:37 and SEQ ID NO:38, as well asamino acid SEQ ID NO:39. The top line in Table 1 corresponds to SEQ IDNO:37, the middle line corresponds to SEQ ID NO:38, and the bottom linecorresponds to the amino acid SEQ ID NO:39. TABLE 1GCGCAGCGGCAACAGCGCTGACGGTACGGACCCACAGTGTACTGAAGGGTATAGTGCAAC----------+--------+---------+---------+---------+---------+CGCGTCGCCGTTGTCGCGACTGCCATGCCTGGGTGTCACATGACTTCCCATATCACGTTG  A  A  A  T  A  L  T  V  R  T  H  S  V  L  K  G  I  V  Q  QAGCAGGACAACCTGCTGAGAGCGATACAGGCCCAGCAACACTTGCTGAGGTTATCTGTAT---------+--------+---------+-----+----+---------+---------+TCGTCCTGTTGGACGACTCTCGCTATGTCCGGGTCGTTGTGAACGACTCCAATAGACATA  Q  D  N  L  L  R  A  I  Q  A  Q  Q  H  L  L  R  L  S  V  WGGGGTATTAGACAACTCCGAGCTCGCCTGCAAGCCTTAGAAACCCTTATACAGAATCAGC---------+--------+----------+---------+---------+---------+CCCCATAATCTGTTGAGGCTCGAGCGGACGTTCGGAATCTTTGGGAATATGTCTTAGTCG  G  I  R  Q  L  R  A  R  L  Q  A  L  E  T  L  I  Q  N  Q  QAACGCCTAAACCTAT ---------+-----  195 TTGCGGATTTGGATA   R  L  N  L

EXAMPLE 5

[0062] The found nucleotide sequence from Table 1 was examined forhomologous sequences in the GENEBANK database (Release 72, June 1992)using the GCG computer program (Genetic Computer Group, Inc., WisconsinUSA, Version 7.1, March 1992). Most of the nucleotide sequences ofimmunodeficient viruses of human origin and of isolates from primatesknown by July 1992 are contained in this database.

[0063] The highest homology shown by the nucleotide sequence from Table1, of 66%, is to a chimpanzee isolate. The highest homology shown by theinvestigated DNA sequence from MVP-5180/91 (SEQ ID NO:56) to HIV-1isolates is 64%. The DNA from Table 1 is 56% homologous to HIV-2isolates. Apart from the chimpanzee isolate sequence, the best homologybetween the nucleotide sequence from Table 1 (SEQ ID NO:37; SEQ IDNO:38) and segments of DNA from primate isolates (SIV: simianimmunodeficiency virus) is found with a DNA sequence encoding a part ofthe coat protein region from the SIV isolate (African long-tailedmonkey) TYO-1. The homology is 61.5%.

EXAMPLE 6

[0064] The found amino acid sequence from Table 1 (SEQ ID NO:39) wasexamined for homologous sequences in the SWISSPROT protein database(Release 22, June 1992) using the GCG computer program. Most of theprotein sequences of immunodeficiency viruses of human origin and ofisolates from primates known by June 1992 are contained in thisdatabase.

[0065] The highest homology shown by the amino acid sequence from Table1 (SEQ ID NO:39), of 62.5%, is to a segment of coat protein from theabovementioned chimpanzee isolate. The best homology among HIV-1 coatproteins to the amino acid sequence from Table 1 (SEQ ID NO:39) is foundin the isolate HIV-1 Mal. The homology is 59%. The highest homology ofthe amino acid sequence from Table 1 (SEQ ID NO:39) to HIV-2 coatproteins is 52% (isolate HIV-2 Rod). Since HIV-1 and HIV-2 isolates,themselves, are at most only 64% identical in the corresponding proteinsegment, the MVP-5180/91 (SEQ ID NO:56) isolate appears to be an HIVvariant which clearly differs structurally from HIV-1 and HIV-2 and thusrepresents an example of an independent group of HIV viruses.

[0066] The amino acid sequence of the amplified region of DNA (Table 1;SEQ ID NO:39) from the HIV isolate MVP-5180/91 (SEQ ID NO:56) overlapsan immunodiagnostically important region of the coat protein gp 41 fromHIV-1 (amino acids 584-618*) (Table 2, which includes SEQ ID NO:61 asthe top line and SEQ ID NO:63 as the bottom line) (Gnann et al., J. Inf.Dis. 156: 261-267, 1987; Norrby et al., Nature, 329: 248-250, 1987).

[0067] Corresponding amino acid regions from the coat proteins of HIV-2and SIV are likewise immunodiagnostically conserved (Gnann et al.,Science, pp. 1346-1349, 1987). Thus, peptides from this coat proteinregion of HIV-1 and HIV-2 are employed as solid-phase antigens in manycommercially available HIV-1/2 antibody screening tests. Approximately99% of the anti-HIV-1 and anti-HIV-2 positive sera can be identified bythem.

[0068] The amino acid region of the MVP-5180/91 coat protein (Table 1)could be of serodiagnostic importance owing to the overlap with theimmunodiagnostically important region from gp 41. This would be the caseparticularly if antisera from HIV-infected patients failed to reactpositively with any of the commercially available antibody screeningtests. In these cases, the infection could be with a virus which wasclosely related to MVP-5180/91 (SEQ ID NO:56). TABLE 2 (includes SEQ IDNO:61 as the top line and SEQ ID NO:63 as the bottom line)         .         .         .         .........RILAVERYLKDQQLLGIWGCSGKLICTTAVPWNAS         |:|:| .:.:|| |.:WGIRQLRARLQALETLIQNQQRLNL..................

EXAMPLE 7

[0069] DNA isolation, amplification and structural characterization ofgenome segments from the HIV isolate MVP-5180/91 (SEQ ID NO:56)(encoding gp 41).

[0070] Genomic DNA from MVP-5180/91 (SEQ ID NO:56)-infected HUT 78 cellswas isolated as described.

[0071] In order to characterize genomic regions of the isolateMVP-5180/91, PCR (polymerase chain reaction) experiments were carriedout using primer pairs from the gp 41 coat protein region. PCR (Saiki etal., Science 239: 487-491, 1988) and inverse PCR (Triglia et al., Nucl.Acids, Res. 16: 8186, 1988) were carried out with the followingmodifications:

[0072] 1. PCR

[0073] For the amplification of HIV-specific DNA regions, 5 μl (218μg/ml) of genomic DNA from MVP-5180/91 infected HUT 78 cells werepipetted into a 100 μl reaction mixture (0.25 mM dNTP, in each case 1 μmprimer 163env (SEQ ID NO:40) and primer envend (SEQ ID NO:41), 10 mMTris HCl, pH 8.3, 50 mM KCl, 1.5 mM MgCl₂, 0.001% gelatin, 2.5 units ofTaq polymerase (Perkin Elmer)), and amplification was then carried outin accordance with the following temperature program: 1. initialdenaturation: 3 min. 95° C., 2. amplification: 90 sec. 94° C., 60 sec.56° C., 90 sec. 72° C. (30 cycles).

[0074] 2. Inverse PCR

[0075] The 5′ region of gp 41 (N terminus) and the 3′ sequence of gp 120were amplified by means of “inverse PCR”. For this, 100 μl of a genomicDNA preparation (218 μg/ml) from MVP-5180/91-infected HUT 78 cells weredigested at 37° C. for 1 hour in a final volume of 200 μl using 10 unitsof the restriction endonuclease Sau3a. The DNA was subsequentlyextracted with phenol and then precipitated using sodium acetate (finalconcentration 300 mM) and 2.5 volumes of ethanol, with storage at −70°C. for 10 min, and then centrifuged down in an Eppendorf centrifuge; thepellet was then dried and resuspended in 890 μl of distilled water.Following addition of 100 μl of ligase buffer (50 mM Tris HCl, pH 7.8,10 mM MgCl₂₁ 10 mM DTT, 1 mM ATP, 25 μg/ml bovine serum albumin) and 10μl of T4 DNA ligase (from Boehringer, Mannheim), the DNA fragments wereligated at room temperature for 3 hours and then extracted with phenolonce again and precipitated with sodium acetate and ethanol as above.After centrifuging down and drying, the DNA was resuspended in 40 μl ofdistilled water and digested for 1 hour with 10 units of the restrictionendonuclease SacI (from Boehringer, Mannheim). 5 μl of this mixture werethen employed in a PCR experiment as described under “1. PCR”. Theprimers 168i (SEQ ID NO:42) and 169i (SEQ ID NO:43) were used for theinverse PCR in place of primers 163env (SEQ ID NO:40) and envend (SEQ IDNO:41).

[0076] The primers 163env (SEQ ID NO:40), 168i (SEQ ID NO:42) and 169i(SEQ ID NO:43) were selected from that part of the sequence of the HIVisolate MVP-5180 (SEQ ID NO:56) which had already been elucidated(Example 4).

[0077] The primers used for the PCR/inverse PCR and the nucleotidesequencing were synthesized on a Biosearch 8750 oligonucleotidesynthesizer, with the primers having the following sequences: Primer163env (SEQ ID NO:40) 5′ CAG AAT CAG CAA CGC CTA AAC C 3′ Primer envend(SEQ ID NO:41): 5′ GCC CTG TCT TAT TCT TCT AGG 3′ (position from HIV-1isolate BH10: bases 8129-8109) Primer 168i (SEQ ID NO:42): 5′ CCC TGCAAG CCT TAG AAA CC 3′ Primer 169i (SEQ ID NO:43) 5′ GCA CTA TAC CCT TCAGTA CAC TG 3′

[0078] The amplified DNA was fractionated on a 3% “Nusieve” agarose gel(from Biozyme) and the amplified fragment was then cut out and an equalvolume of buffer (1*TBE (0.09 M Tris borate, 0.002 M EDTA, pH 8.0)) wasadded to it. After incubating the DNA/agarose mixture at 70° C. for 10minutes, and subsequent phenol extraction, the DNA was precipitated fromthe aqueous phase by adding {fraction (1/10)} vol of 3 M NaAc, pH 5.5,and 2 vol of ethanol, and storing at −20° C. for 15′, and then pelletedin an Eppendorf centrifuge (13,000 rpm, 10′, 4° C.). The pelleted DNAwas dried and then taken up in water and sequenced by the method ofSanger (F. Sanger, Proc. Natl. Acad. Sci., 74: 5463, 1977) followingphotometric determination of the DNA concentration at 260 nm in aspectrophotometer (from Beckman). Instead of sequencing with Klenow DNApolymerase, the sequencing reaction was carried out using a kit fromApplied Biosystems (“Taq dye deoxy terminator cycle sequencing”, orderNo.: 401150). Primer 163env (SEQ ID NO:40) or primer envend (SEQ IDNO:41) (in each case 1 μM) was employed as the primer in separatesequencing reactions. The amplified DNA from the inverse PCR experimentwas sequenced using primers 168i (SEQ ID NO:42) and 169i (SEQ ID NO:43).The sequencing reaction was analysed on an Applied Biosystems 373A DNAsequencing apparatus in accordance with the instructions of theapparatus manufacturer.

[0079] The nucleotide sequence of the amplified DNA region, and theamino acid sequence deduced from it, are presented in Table 3. Table 3includes DNA sequences SEQ ID NO:44 and SEQ ID NO:45, as well as aminoacid sequence SEQ ID NO:46. In Table 3, the top line corresponds to SEQID NO:44, the middle line corresponds to SEQ ID NO:45, and the bottomline represents amino acid sequence SEQ ID NO:46. TABLE 3AAATGTCAAGACCAATAATAAACATTCACACCCCTCACAGGGAAAAAAGAGCAGTAGGAT---------+---------+---------+---------+---------+---------+ 60TTTACAGTTCTGGTTATTATTTGTAAGTGTGGGGAGTGTCCCTTTTTTCTCGTCATCCTA  M  S  R  P  I  I  N  I  H  T  P  H  R  E  K  R  |A  V  G  L                                     gp120            gp41TGGGAATGCTATTCTTGGGGGTGCTAAGTGCAGCAGGTAGCACTATGGGCGCAGCGGCAA---------+---------+---------+---------+---------+---------+ 120ACCCTTACGATAAGAACCCCCACGATTCACGTCGTCCATCGTGATACCCGCGTCGCCGTT  G  M  L  F  L  G  V  L  S  A  A  G  S  T  M  G  A  A  A  TCAGCGCTGACGGTACGGACCCACAGTGTACTGAAGGGTATAGTGCAACAGCAGGACAACC---------+---------+---------+---------+---------+---------+ 180GTCGCGACTGCCATGCCTGGGTGTCACATGACTTCCCATATCACGTTGTCGTCCTGTTGG  A  L  T  V  R  T  H  S  V  L  K  G  I  V  Q  Q  Q  D  N  LTGCTGAGAGCGATACAGGCCCAGCAACACTTGCTGAGGTTATCTGTATGGGGTATTAGAC---------+---------+---------+---------+---------+---------+ 240ACGACTCTCGCTATGTCCGGGTCGTTGTGAACGACTCCAATAGACATACCCCATAATCTG  L  R  A  I  Q  A  Q  Q  H  L  L  R  L  S  V  W  G  I  R  QAACTCCGAGCTCGCCTGCAAGCCTTAGAAACCCTTATACAGAATCAGCAACGCCTAAACC---------+---------+---------+---------+---------+---------+ 300TTGAGGCTCGAGCGGACGTTCGGAATCTTTGGGAATATGTCTTAGTCGTTGCGGATTTGG  L  R  A  R  L  Q  A  L  E  T  L  I  Q  N  Q  Q  R  L  N  LTATGGGGCTGTAAAGGAAAACTAATCTGTTACACATCAGTAAAATGGAACACATCATGGT---------+---------+---------+---------+---------+---------+ 360ATACCCCGACATTTCCTTTTGATTAGACAATGTGTAGTCATTTTACCTTGTGTAGTACCA  W  G  C  K  G  K  L  I  C  Y  Y  S  V  K  W  N  T  S  W  SCAGGAGGATATAATGATGACAGTATTTGGGACAACCTTACATGGCAGCAATGGGACCAAC---------+---------+---------+---------+---------+---------α 420GTCCTCCTATATTACTACTGTCATAAACCCTGTTGGAATGTACCGTCGTTACCCTGGTTG  G  G  Y  N  D  D  S  I  W  D  N  L  T  W  Q  Q  W  D  Q  HACATAAACAATGTAAGCTCCATTATATATGATGAAATACAAGCAGCACAAGACCAACAGG---------+---------+---------+---------+---------+---------+ 480TGTATTTGTTACATTCGAGGTAATATATACTACTTTATGTTCGTCGTGTTCTGGTTGTCC  I  N  N  V  S  S  I  I  Y  D  E  I  Q  A  A  Q  D  Q  Q  EAAAAGAATGTAAAAGCATTGTTGGAGCTAGATGAATGGGCCTCTCTTTGGAATTGGTTTG---------+---------+---------+---------+---------+---------+ 540TTTTCTTACATTTTCGTAACAACCTCGATCTACTTACCCGGAGAGAAACCTTAACCAAAC  K  N  V  K  A  L  L  E  L  D  E  W  A  S  L  W  N  W  F  DACATAACTAAATGGTTGTGGTATATAAAAATAGCTATAATCATAGTGGGAGCACTAATAG---------+---------+---------+---------+---------+---------+ 600TGTATTGATTTACCAACACCATATATTTTTATCGATATTAGTATCACCCTCGTGATTATC  I  T  K  W  L  W  Y  I  K  I  A  I  I  I  V  G  A  L  I  GGTATAAGAGTTATCATGATAGTACTTAATCTAGTGAAGAACATTAGGCAGGGATATCAAC---------+---------+---------+---------+---------+---------+ 660CATATTCTCAATAGTACTATCATGAATTAGATCACTTCTTGTAATCCGTCCCTATAGTTG  I  R  V  I  M  I  V  L  N  L  V  K  N  I  R  Q  G  Y  Q  PCCCTCTCGTTGCAGATCCCTGTCCCACACCGGCAGGAAGCAGAAACGCCAGGAAGAACAG---------+---------+---------+---------+---------+---------+ 720GGGAGAGCAACGTCTAGGGACAGGGTGTGGCCGTCCTTCGTCTTTGCGGTCCTTCTTGTC  L  S  L  Q  I  P  V  P  H  R  Q  E  A  E  T  P  G  R  T  GGAGAAGAAGGTGGAGAAGGAGACAGGCCCAAGTGGACAGCCTTGCCACCAGGATTCTTGC---------+---------+---------+---------+---------+---------+ 780CTCTTCTTCCACCTCTTCCTCTGTCCGGGTTCACCTGTCGGAACGGTGGTCCTAAGAACG  E  E  G  G  E  G  D  R  P  K  W  T  A  L  P  P  G  F  L  QAACAGTTGTACACGGATCTCAGGACAATAATCTTGTGGACTTACCACCTCTTGAGCAACT---------+---------+---------+---------+---------+---------+ 840TTGTCAACATGTGCCTAGAGTCCTGTTATTAGAACACCTGAATGGTGGAGAACTCGTTGA  Q  L  Y  T  D  L  R  T  I  I  L  W  T  Y  H  L  L  S  N  LTAATATCAGGGATCCGGAGGCTGATCGACTACCTGGGACTGGGACTGTGGATCCTGGGAC---------+---------+---------+---------+---------+---------+ 900ATTATAGTCCCTAGGCCTCCGACTAGCTGATGGACCCTGACCCTGACACCTAGGACCCTG  I  S  G  I  R  R  L  I  D  Y  L  G  L  G  L  W  I  L  G  QAAAAGACAATTGAAGCTTGTAGACTTTGTGGAGCTGTAATGCAATATTGGCTACAAGAAT---------+---------+---------+---------+---------+---------+ 960TTTTCTGTTAACTTCGAACATCTGAAACACCTCGACATTACGTTATAACCGATGTTCTTA  K  T  I  E  A  C  R  L  C  G  A  V  M  Q  Y  W  L  Q  E  LTGAAAAATAGTGCTACAAACCTGCTTGATACTATTGCAGTGTCAGTTGCCAATTGGACTG---------+---------+---------+---------+---------+---------+ 1020ACTTTTTATCACGATGTTTGGACGAACTATGATAACGTCACAGTCAACGGTTAACCTGAC  K  N  S  A  T  N  L  L  D  T  I  A  V  S  V  A  N  W  T  DACGGCATCATCTTAGGTCTACAAAGAATAGGACAAGG---------+---------+---------+------- 1057TGCCGTAGTAGAATCCAGATGTTTCTTATCCTGTTCC   G  I  I  L  G  L  Q  R  I  G  Q

EXAMPLE 8

[0080] The found nucleotide sequence from Table 3 (SEQ ID NO:44; SEQ IDNO:45) was examined for homologous sequences in the GENEBANK database(Release 72, June 1992) using the GCG computer program (Genetic ComputerGroup, Inc. Wisconsin USA, version 7.1, March 1992). Most of thenucleotide sequences of immunodeficiency viruses of human origin and ofisolates from primates known by July 1992 are contained in thisdatabase.

[0081] The highest homology of the nucleotide sequence from Table 3 (SEQID NO:44; SEQ ID NO:45) to an HIV-1 isolate is 62%. The DNA from Table 3is 50% homologous to HIV-2 isolates.

[0082] The amino acid sequence deduced from the nucleotide sequence fromTable 3 (SEQ ID NO:46) was examined for homologous sequences in theSWISSPROT protein database (Release 22, June 1992) using the GCGcomputer program. Most of the protein sequences of immunodeficiencyviruses of human origin and of isolates from primates known by June 1992are contained in this database.

[0083] At best, the amino acid sequence from Table 3 (SEQ ID NO:46) is54% homologous to the corresponding coat protein segment from achimpanzee isolate CIV (SIVcpz) and 54.5% homologous to the HIV-1isolate Mal. At best, the amino acid sequence from Table 3 (SEQ IDNO:46) is 34% homologous to HIV-2 coat proteins (isolate HIV-2 D194).

[0084] If, by contrast, the gp 41 amino acid sequence of HIV-1 iscompared with the HIV-1 gp 41 sequence present in the SWISSPROTdatabase, the highest homology is, as expected, almost 100%, and thelowest 78%.

[0085] These clear structural differences between the sequence regionfrom Table 3 and the corresponding segment from HIV-1 and HIV-2 suggestthat isolate MVP-5180/91 (SEQ ID NO:56) is an HIV variant which clearlydiffers structurally from HIV-1 and HIV-2. It is possible thatMVP-5180/91 (SEQ ID NO:56) should be assigned to a separate group of HIVviruses which differ from HIV-1 and HIV-2.

[0086] The peptide from amino acid 584 to amino acid 618 of the HIV-1coat protein region (SEQ ID NO:61) is of particular serodiagnosticinterest (numbering in accordance with Wain Hobson et al., Cell 40:9-17, 1985; Gnann et al., J. Inf. Dis. 156: 261-267, 1987; Norrby etal., Nature, 329: 248-250, 1987). Corresponding amino acid regions fromthe coat proteins of HIV-2 and SIV are likewise immunodiagnosticallyconserved (Gnann et al., Science, pp. 1346-1349, 1987). Thus, peptidesfrom this coat protein region of HIV-1 and HIV-2 are employed assolid-phase antigens in many commercially available HIV-1/2 antibodyscreening tests. Using them, approximately 99% of the anti-HIV-1 andanti-HIV-2-positive sera can be identified.

[0087] The corresponding amino acid region of the MVP-5180/91 coatprotein (Table 4), as well as the whole gp 41 of this isolate, could beof serodiagnostic importance, particularly if antisera from HIV-infectedpatients either did not react at all or only reacted weakly incommercially available antibody screening tests. In these cases, theinfection could be due to a virus which is closely related toMVP-5180/91 (SEQ ID NO:56).

[0088] Table 4 includes SEQ ID NO:61, which is designated as line 1, andalso highlights in line 2 the points of difference from the amino acidsequence designated SEQ ID NO:62. μmino acid sequence SEQ ID NO:62appears in full following Table 4. TABLE 4 1RILAVERYLKDQQLLGIWOCSGKLICTTAVPWNAS 2  LQ L TLIQN R NL    K    Y S K T

[0089] The peptide, which was found with the aid of information derivingfrom MVP-5180, thus has the amino acid sequence (SEQ ID NO:62):RLQALETLIQNQQRLNLWGCKGKLICYTSVKWNTS.

[0090] The present invention therefore relates to peptides which can beprepared recombinantly or synthetically and have the sequence indicatedabove, or a constituent sequence thereof, the constituent sequenceshaving at least 6 consecutive amino acids, preferably 9 and particularlypreferably 12 consecutive amino acids.

EXAMPLE 9:

[0091] Cloning of the whole genome of the HIV isolate MVP-5180 (SEQ IDNO:56)

[0092] a) Preparation of a genomic library

[0093] Genomic DNA from MVP-5180-infected HUT 78 cells was isolated asdescribed.

[0094] 300 μg of this DNA were incubated for 45 min in a volume of 770μl together with 0.24 U of the restriction enzyme Sau3A. The DNA, whichwas only partially cut in this incubation, was subsequentlysize-fractionated on a 0.7% agarose gel (low melting agarose, Nusieve)and fragments of between 10 and 21 kb were cut out. The agarose wasmelted at 70° C. for 10 min and the same volume of buffer (1*TBE, 0.2 MNaCl) was then added to it. Subsequently, after having extracted twicewith phenol and once with chloroform, the DNA was precipitated by adding1/10 vol. of 3 M sodium acetate solution (pH 5.9) and 2.5 vol. ofethanol, and storing at −700° C. for 10 min. The precipitated DNA wascentrifuged down and dried and then dissolved in water at aconcentration of 1 μg/μl.

[0095] The yield of size-fractionated DNA was about 60 μg. 5 μg of thisDNA were incubated at 37° C. for 20 min in an appropriate buffertogether with 1 U of alkaline phosphatase. In this way, the risk ofmultiple insertions of size-fractionated DNA was reduced by eliminatingthe 5′-terminal phosphate radical. The phosphatase treatment was stoppedby extracting with phenol and the DNA was precipitated as above and thenligated at 15° C. for 12 hours together with 1 μg of the vector (2 DASH,BamHI-cut, Stratagene No.: 247611) in a total volume of 6 μl using 2Weiss units of Lambda T4 ligase. Following completed ligation, the DNAwas packaged into phage coats using a packaging kit (Gigapack II Gold,Stratagene No.: 247611) precisely in accordance with the manufacturer'sinstructions.

[0096] b) Radioactive labeling of the DNA probe.

[0097] The “random-primed DNA labeling kit” from Boehringer Mannheim(No.: 713 023) was employed for the labeling. The PCR product waslabeled which was obtained as described in Example 3 using the primerssk68 (SEQ ID NO:21) and envb (SEQ ID NO:20). 1 μg of this DNA wasdenatured by 2*5 min of boiling and subsequent cooling in ice water. 50mCi [a- ³²p]-dCTP (NEN, No.: NEX-053H) were added for the labeling.Other ingredients were added by pipette in accordance with themanufacturer's instructions. Following a 30 min incubation at 37° C.,the DNA, which was now radioactively labeled, was precipitated.

[0098] c) Screening the phage library

[0099] 20,000 pfu (plaque-forming units) of the library in 100 μl of SMbuffer (5.8 g of NaCl, 2 g of MgSO₄, 50 ml of 1 M Tris, pH 7.5, and 5 mlof a 2% gelatin solution, dissolved in 1 l of H₂O ) were added to 200 μlof a culture (strain SRB(P2) [Stratagene, No.: 247611] in LB medium,which contained 10 mM MgSO₄ and 0.2% maltose) which had been grown at30° C. overnight; the phages were adsorbed to the bacteria at 37° C. for20 min and 7.5 ml of top agarose, which had been cooled to 55° C., wasthen mixed in and the whole sample was distributed on a pre-warmed LBagar plate of 14 cm diameter. The plaques achieved confluence afterabout 8 hours. After that, nitrocellulose filters were laid on theplates for a few minutes and were marked asymmetrically. After havingbeen carefully lifted from the plates, the filters were denatured for 2min (0.5 M NaOH, 1.5 M NaCl) and then neutralized for 5 min (0.5 M Tris,pH 8, 1.5 M NaCl). The filters were subsequently baked at 80° C. for 60min and could then be hybridized to the probe. For the prehybridization,the filters were incubated at 42° C. for 2-3 h, while shaking, in 15 mlof hybridization solution (50% formamide, 0.5% SDS, 5*SSPE, 5*Denhardt'ssolution and 0.1 mg/ml salmon sperm DNA) per filter. The [³²]-labeledDNA probes were denatured at 100° C. for 2-5 min and then cooled on ice;they were then added to the prehybridization solution and hybridizationwas carried out at 42° C. for 12 hours. Subsequently, the filters werewashed at 60° C., firstly with 2*SSC/0.1% SDS and then with 0.2*SSC/0.1%SDS. After the filters had been dried, hybridization signals weredetected using the X-ray film X-OMAT™AR (Kodak).

[0100] Following elution in SM buffer, those plaques to which it waspossible to assign a signal were individually separated in furtherdilution steps.

[0101] It was possible to identify the clone described below followingscreening of 2*10⁶ plaques.

[0102] d) Isolation of the phage DNA and subcloning

[0103] An overnight culture of the host strain SRB (P2) was infectedwith 10 11 of a phage eluate in SM buffer such that the cultureinitially grew densely but then lysed after about 6-8 h. Cell remnantswere separated off from the lysed culture by centrifuging it twice at9,000 g for 10 min. Subsequently, the phage were pelleted bycentrifugation (35,000 g, 1 h), and then taken up in 700 μl of 10 mMMgSO₄ and extracted with phenol until a protein interface could nolonger be seen. The phage DNA was then precipitated and cleaved with therestriction enzyme EcoRI, and the resulting EcoRI fragments weresubcloned into the vector Bluescript, KS⁻ (Stratagene, No.: 212208). Inall, 4 clones were obtained: Plasmid Beginning¹ End¹ pSP1 1 1785 pSP21786 5833 pSP3 5834 7415 pSP4 7660 9793

[0104] The missing section between bases 7416 and 7659 was obtained byPCR using the primers 157 (CCA TAA TAT TCA GCA GAA CTA G) (SEQ ID NO:64)and 226 (GCT GAT TCT GTA TAA GGG) (SEQ ID NO:65). The phage DNA of theclone was used as the DNA template. The conditions for the PCR were: 1.)initial denaturation: 94° C., 3 min, 2.) amplification: 1.5 min 94° C.,1 min 56° C. and 1 min 72° C. for 30 cycles.

[0105] The DNA was sequenced as described in Example 4. Both the strandand the antistrand of the total genome were sequenced. In the case ofeach site for EcoRI cleavage, PCR employing phage DNA of the clone asthe DNA template was used to verify that there was indeed only the oneEcoRI cleavage site at each subclone transition point.

[0106] Table 5: The position of the genes for the virus proteins GAG,POL and ENV in the full sequence of MVP-5180 Gene Start¹ Stop¹ GAG 8172310 POL 2073 5153 ENV 6260 8887

EXAMPLE 10

[0107] Delimitation of the full sequence of MVP-5180/91 (SEQ ID NO:56)from other HIV-1 isolates

[0108] The databanks Genbank, Release 75 of 2.93, EMBL 33 of 12.92, andSwissprot 24 of 1.93 provided the basis for the following sequencecomparisons. Comparisons of homology were carried out using the GCGsoftware (version 7.2, 10.92. from the Genetics Computer Group,Wisconsin).

[0109] Initially, the sequences of GAG, POL and ENV were compared withthe database at the amino acid level using the “Wordsearch” program. The50 best homologs were in each case compared with each other using the“Pileup” program. From this, it clearly emerges that MVP-5180/91 (SEQ IDNO:56) belongs in the HIV-1 genealogical tree but branches off from itat a very early stage, even prior to the chimpanzee virus SIVcpz, andthus represents a novel HIV-1 subfamily. In order to obtain numericalvalues for the homologies, MVP-5180 (SEQ ID NO:56) was compared with theHIV-1, HIV-2 and SIV sequences which in each case showed the best fit,and in addition with the SIVcpz sequence, using the “Gap” program. TABLE6 Homology values for the amino acid sequences of GAG, POL and ENV ofthe MVP-5180/91 isolate GAG SIVcpz 70.2% HIV1u² 69.9% HIV2d³ 53.6%SIV1a⁴ 55.1% 83.6% 81.2% 71.3% 71.3% POL SIVcpz 78.0% HIV1u² 76.1%HIV2d³ 57.2% SIVgb⁵ 57.7% 88.0% 86.8% 71.9% 74.6% ENV SIVcpz 53.4%HIV1h¹ 50.9% HIV2d³ 34.4% SIVat⁶ 34.4% 67.1% 67.2% 58.7% 57.8%

[0110] The upper numerical value expresses the identity and the lowervalue the similarity of the two sequences.

[0111] In addition to this, the database was searched at the nucleotidelevel using “Wordsearch” and “Gap”. The homology values for the bestmatches in each- case are compiled in Table 7. TABLE 7 [126] Homologyvalues for the nucleotide sequence of MVP-5180/91 HIV1 HIV2 gag HIVelicg70.24% HIV2bihz 60.0% pol HIVmal 75.0% HIV2cam2 62.9% env HIVsimi8459.7% HIV2gha 49.8

EXAMPLE 11

[0112] Description of the PCR amplification, cloning and sequencing ofthe gag gene of the HIV 5180 isolate.

[0113] In order to depict the spontaneous mutations arising during thecourse of virus multiplication, a part of the viral genome was clonedusing the PCR technique and the DNA sequence thus obtained was comparedwith the sequence according to FIG. 4 (SEQ ID NO:56).

[0114] The gag sequence was cloned in an overlapping manner from the LTR(long terminal repeat, LTRl primer) of the left end of the MVP-5180genome through into the pol gene (polymerase gene, pol3.5i primer). Thecloning strategy is depicted schematically in FIG. 5.

[0115] The PCR reactions were carried out using the DNA primers givenbelow, whose sequences were derived from the HIV-1 consensus sequence.The sequencings were carried out using the dideoxy chain terminationmethod.

[0116] The sequence encoding the MVP-5180 gag gene extends fromnucleotide 817 (A of the ATG start codon) to nucleotide 2300 (A of thelast codon). LTR1 (SEQ ID NO:47): 5′ -CTA GCA GTG GCG CCC GAA CAG G-3′gag3.5 (SEQ ID NO:48): 5′ -AAT GAG GAA GCU GCA GAU TGG GA-3′       (U= A/T) gag3.5i (SEQ ID NO:49): 5′ -TCC CAU TCT GCU GCT TCC TCA TT-3′      (U = A/T) gag5 (SEQ ID NO:50): 5′-CCA AGG GGA AGT GAC ATA GCA GGAAC-3′ gag959 (SEQ ID NO:51): 5′-CGT TGT TCA GAA TTC AAA CCC-3′ gag11(SEQ ID NO:52): 5′-TCC CTA AAA AAT TAG CCT GTC-3′ po13.5i (SEQ IDNO:53): 5′-AAA CCT CCA ATT CCC CCT A-3′

[0117] The DNA sequence obtained using the PCR technique was comparedwith the DNA sequence presented in FIG. 4 (SEQ ID NO:56). A comparisonof the two DNA sequences is presented in FIG. 6. FIG. 6 includes SEQ IDNO:57, which corresponds to FIG. 4 (SEQ ID NO:56) and SEQ ID NO:58,which corresponds to the DNA sequence obtained using the PCR technique.This showed that about 2% of the nucleotides differ from each other,although the virus is the same in the two cases. In FIG. 6, the upperline in each case represents the DNA sequence which is presented in FIG.4 (SEQ ID NO:56) and the lower line represents the DNA sequence obtainedusing the PCR technique.

[0118] In addition, the amino acid sequence of the gag protein,elucidated using the PCR technique, was compared with the amino acidsequence of the corresponding protein deduced from FIG. 4 (SEQ IDNO:59). This showed an amino acid difference of about 2.2%. Thecomparison is presented in FIG. 7, the lower line in each caserepresenting the amino acid sequence which was deduced from the sequenceobtained using the PCR technique. FIG. 7 includes amino acid SEQ IDNO:59, which was elucidated in accordance with FIG. 4 (SEQ ID NO:56),and the amino acid sequence (SEQ ID NO:60) derived using the PCRtechnique.

EXAMPLE 12

[0119] The sequence of the virus MVP-5180 (SEQ ID NO:56) according tothe invention was compared with the consensus sequences of HIV-1 andHIV-2, and with the sequence of ANT-70 (WO 89/12094), insofar as thiswas known.

[0120] In this connection, the following results were obtained: TABLE 8[138] Deviating Number of the % homology Gene locus nucleotidesnucleotides (approximated) LTR 207 630 HIV-1 67% 308 HIV-2 51% 115 ANT70 82% GAG 448 1501 HIV-1 70% 570 HIV-2 62% POL 763 3010 HIV-1 74% 1011HIV-2 66% VIF 183 578 HIV-1 68% 338 HIV-2 42% ENV 1196 2534 HIV-1 53%1289 HIV-2 49% NEF 285 621 HIV-1 54% 342 HIV-2 45% total 3082 8874 HIV-165% 3858 HIV-2 56%

[0121] In the above table, “HIV-1” denotes consensus sequences of HIV-1viruses; “HIV-2” denotes consensus sequences of HIV-2 viruses; ANT-70denotes the partial sequence of a virus designated HIV-3 and disclosedin WO 89/12094.

[0122] The present invention therefore relates to viruses, DNA sequencesand amino acid sequences, and constituent sequences thereof, whichpossess such a degree of homology with the sequence presented in FIG. 4(SEQ ID NO:56), based on the gene loci, that at most the fractions givenin Table 9, expressed in % values, are different. TABLE 9 [141] Homologybased on gene loci, expressed as maximum differences ParticularlyPreferred preferred Gene locus Differences differences differences LTR17% 15% 10% GAG 29% 28% 14% POL 25% 24% 12% VIF 31% 30% 15% ENV 46% 45%22% NEF 16% 12% 10%

[0123] The homology values in % given in Table 9 mean that, whencomparing the sequence according to FIG. 4 (SEQ ID NO:56) with asequence of another virus, at most a fraction of the sequencecorresponding to the abovementioned percentage values may be different.

EXAMPLE 13

[0124] V3 Loop

[0125] This loop is the main neutralizing region in HIV and theimmunological specificities of the region are documented in summary formin FIG. 8. This is a copy from a work by Peter Nara (1990) from AIDS.The amino acid sequence of the V3 loop is shown diagrammatically and iscompared with the IIIB virus, now LAI, and the first HIV-2 isolate(ROD). Individual amino acids are conserved at the cystine bridge.Whereas the crown of HIV-1 is GPGR or GPGQ and that of HIV-2 is GHVF,the crown of MVP-5180/91 (SEQ ID NO:56) is formed from the amino acidsGPMR. The motif with methionine has not previously been described and,emphasizes the individuality of MVP-5180/91 (SEQ ID NO:56).

[0126] After having determined the nucleotide sequence of the virus theV3-loop-region was amplified using the PCR-technique by using suitableprimers. Some mutations have been observed, especially a change of themethionine codon (ATG) to the leucine codon (CTG).

[0127] In the following the amino acid sequence derived from the clonednucleic acid is compared with a sequence obtained after amplificationwith the help of PCR technology. MvP 5180 (cloned) (SEQ ID NO:54):CIREGIAEVQDIYTGPMRWRSMTLKRSNNTSPRSRVAYC MvP 5180 (PCT technique) (SEQ IDNO:55) CIREGIAEVQDLHTGPLRWRSMTLKKSSNSHTQPRSKVAYC

EXAMPLE 14

[0128] In order to demonstrate that even those sera which cannot beidentified in a normal HIV-1+2 screening test can be proved to beHIV-1-positive with the aid of the virus MVP-5180 (SEQ ID NO:56)according to the invention, or antigens derived therefrom, various serafrom patients from the Cameroons were examined in the EIA test.

[0129] 156 anti-HIV-1-positive sera were examined in a study carried outin the Cameroons. Substantial, diagnostically relevant differences wereobserved in two of these sera. The extinctions which were measured aregiven in Table 10 below. CAM-A and CAM-B denote the sera of differentpatients. TABLE 10 [150] Patient sera MVP-5180-EIA HIV-1 + HIV-2 EIACAM-A 2.886 1.623 CAM-B 1.102 0.386

[0130] The cutoff for both tests was 0.300.

[0131] In a further study on 47 anti-HIV-1-positive sera from theCameroons, two sera were of particular note. One of these (93-1000)derives from a patient showing relatively few symptoms and the other(93-1001) from a patient suffering from AIDS. The extinction values forthe two EIA tests are compared in Table 11 below: TABLE 11 [152] Patientsera MVP-5180-EIA HIV-1 + HIV-2 EIA 93-1000 >2.5 1.495 93-1001 0.6920.314

[0132] The cutoff was 0.3 in this case as well. The extinction valuesfor patient 93-1001 demonstrate that the normal HIV-1+HIV-2 EIA can failwhereas clear detection is possible the antigen according to theinvention is employed.

1 67 1 18 DNA Artificial Sequence Description of Artificial Sequenceprimer 1 ctactagtac ccttcagg 18 2 21 DNA Artificial Sequence Descriptionof Artificial Sequence primer 2 cggtctacat agtctctaaa g 21 3 21 DNAArtificial Sequence Description of Artificial Sequence primer 3ccacctatcc cagtaggaga a 21 4 30 DNA Artificial Sequence Description ofArtificial Sequence primer 4 cctttggtcc ttgtcttatg tccagaatgc 30 5 25DNA Artificial Sequence Description of Artificial Sequence primer 5tgggaagttc aattaggaat accac 25 6 26 DNA Artificial Sequence Descriptionof Artificial Sequence primer 6 cctacataga aatcatccat gtattg 26 7 19 DNAArtificial Sequence Description of Artificial Sequence primer 7tggatgtggg tgatgcata 19 8 21 DNA Artificial Sequence Description ofArtificial Sequence primer 8 agcacattgt actgatatct a 21 9 22 DNAArtificial Sequence Description of Artificial Sequence primer 9agtgggggga catcaagcag cc 22 10 22 DNA Artificial Sequence Description ofArtificial Sequence primer 10 tgctatgtca cttccccttg gt 22 11 22 DNAArtificial Sequence Description of Artificial Sequence primer 11ccatgcaaat gttaaaagag ac 22 12 19 DNA Artificial Sequence Description ofArtificial Sequence primer 12 ggcctggtgc aataggccc 19 13 20 DNAArtificial Sequence Description of Artificial Sequence primer 13gtgcttccac agggatggaa 20 14 18 DNA Artificial Sequence Description ofArtificial Sequence primer 14 atcatccatg tattgata 18 15 20 DNAArtificial Sequence Description of Artificial Sequence primer 15aatggagcca gtagatccta 20 16 20 DNA Artificial Sequence Description ofArtificial Sequence primer 16 tgtctccgct tcttcctgcc 20 17 20 DNAArtificial Sequence Description of Artificial Sequence primer 17gagccctgga agcatccagg 20 18 20 DNA Artificial Sequence Description ofArtificial Sequence primer 18 ggagatgcct aaggcttttg 20 19 17 DNAArtificial Sequence Description of Artificial Sequence primer 19tgttccttgg gttcttg 17 20 20 DNA Artificial Sequence Description ofArtificial Sequence primer 20 gagttttcca gagcaacccc 20 21 20 DNAArtificial Sequence Description of Artificial Sequence primer 21agcagcagga agcactatgg 20 22 24 DNA Artificial Sequence Description ofArtificial Sequence primer 22 gccccagact gtgagttgca acag 24 23 22 DNAArtificial Sequence Description of Artificial Sequence primer 23gcacagtaca atgtacacat gg 22 24 22 DNA Artificial Sequence Description ofArtificial Sequence primer 24 cagtagaaaa attcccctcc ac 22 25 31 DNAArtificial Sequence Description of Artificial Sequence primer 25tcaggatcca tgggcagtct agcagaagaa g 31 26 42 DNA Artificial SequenceDescription of Artificial Sequence primer 26 atgctcgaga actgcagcatcgattctggg tcccctcctg ag 42 27 40 DNA Artificial Sequence Description ofArtificial Sequence primer 27 cgagaactgc agcatcgatg ctgctcccaagaacccaagg 40 28 21 DNA Artificial Sequence Description of ArtificialSequence primer 28 ggagctgctt gatgccccag a 21 29 22 DNA ArtificialSequence Description of Artificial Sequence primer 29 tgatgacagcatgtcaggga gt 22 30 25 DNA Artificial Sequence Description of ArtificialSequence primer 30 gctgacattt atcacagctg gctac 25 31 27 DNA ArtificialSequence Description of Artificial Sequence primer 31 tatcacctagaactttaaat gcatggg 27 32 22 DNA Artificial Sequence Description ofArtificial Sequence primer 32 agtccctgac atgctgtcat ca 22 33 22 DNAArtificial Sequence Description of Artificial Sequence primer 33gtggagggga atttttctac tg 22 34 24 DNA Artificial Sequence Description ofArtificial Sequence primer 34 cctgctgctc ccaagaaccc aagg 24 35 20 DNAArtificial Sequence Description of Artificial Sequence primer 35agcagcagga agcactatgg 20 36 20 DNA Artificial Sequence Description ofArtificial Sequence primer 36 gagttttcca gagcaacccc 20 37 195 DNA Humanimmunodeficiency virus CDS (3)..(194) 37 gc gca gcg gca aca gcg ctg acggta cgg acc cac agt gta ctg aag 47 Ala Ala Ala Thr Ala Leu Thr Val ArgThr His Ser Val Leu Lys 1 5 10 15 ggt ata gtg caa cag cag gac aac ctgctg aga gcg ata cag gcc cag 95 Gly Ile Val Gln Gln Gln Asp Asn Leu LeuArg Ala Ile Gln Ala Gln 20 25 30 caa cac ttg ctg agg tta tct gta tgg ggtatt aga caa ctc cga gct 143 Gln His Leu Leu Arg Leu Ser Val Trp Gly IleArg Gln Leu Arg Ala 35 40 45 cgc ctg caa gcc tta gaa acc ctt ata cag aatcag caa cgc cta aac 191 Arg Leu Gln Ala Leu Glu Thr Leu Ile Gln Asn GlnGln Arg Leu Asn 50 55 60 cta t 195 Leu 38 195 DNA Human immunodeficiencyvirus 38 ataggtttag gcgttgctga ttctgtataa gggtttctaa ggcttgcaggcgagctcgga 60 gttgtctaat accccataca gataacctca gcaagtgttg ctgggcctgtatcgctctca 120 gcaggttgtc ctgctgttgc actataccct tcagtacact gtgggtccgtaccgtcagcg 180 ctgttgccgc tgcgc 195 39 64 PRT Human immunodeficiencyvirus 39 Ala Ala Ala Thr Ala Leu Thr Val Arg Thr His Ser Val Leu Lys Gly1 5 10 15 Ile Val Gln Gln Gln Asp Asn Leu Leu Arg Ala Ile Gln Ala GlnGln 20 25 30 His Leu Leu Arg Leu Ser Val Trp Gly Ile Arg Gln Leu Arg AlaArg 35 40 45 Leu Gln Ala Leu Glu Thr Leu Ile Gln Asn Gln Gln Arg Leu AsnLeu 50 55 60 40 22 DNA Artificial Sequence Description of ArtificialSequence primer 40 cagaatcagc aacgcctaaa cc 22 41 21 DNA ArtificialSequence Description of Artificial Sequence primer 41 gccctgtcttattcttctag g 21 42 20 DNA Artificial Sequence Description of ArtificialSequence primer 42 gcctgcaagc cttagaaacc 20 43 23 DNA ArtificialSequence Description of Artificial Sequence primer 43 gcactatacccttcagtaca ctg 23 44 1057 DNA Human immunodeficiency virus CDS(3)..(1055) 44 aa atg tca aga cca ata ata aac att cac acc cct cac agggaa aaa 47 Met Ser Arg Pro Ile Ile Asn Ile His Thr Pro His Arg Glu Lys 15 10 15 aga cga gta gga ttg gga atg cta ttc ttg ggg gtg cta agt gca gca95 Arg Arg Val Gly Leu Gly Met Leu Phe Leu Gly Val Leu Ser Ala Ala 20 2530 ggt agc act atg ggc gca gcg gca aca gcg ctg acg gta cgg acc cac 143Gly Ser Thr Met Gly Ala Ala Ala Thr Ala Leu Thr Val Arg Thr His 35 40 45agt gta ctg aag ggt ata gtg caa cag cag gac aac ctg ctg aga gcg 191 SerVal Leu Lys Gly Ile Val Gln Gln Gln Asp Asn Leu Leu Arg Ala 50 55 60 atacag gcc cag caa cac ttg ctg agg tta tct gta tgg ggt att aga 239 Ile GlnAla Gln Gln His Leu Leu Arg Leu Ser Val Trp Gly Ile Arg 65 70 75 caa ctccga gct cgc ctg caa gcc tta gaa acc ctt ata cag aat cag 287 Gln Leu ArgAla Arg Leu Gln Ala Leu Glu Thr Leu Ile Gln Asn Gln 80 85 90 95 caa cgccta aac cta tgg ggc tgt aaa gga aaa cta atc tgt tac aca 335 Gln Arg LeuAsn Leu Trp Gly Cys Lys Gly Lys Leu Ile Cys Tyr Thr 100 105 110 tca gtaaaa tgg aac aca tca tgg tca gga gga tat aat gat gac agt 383 Ser Val LysTrp Asn Thr Ser Trp Ser Gly Gly Tyr Asn Asp Asp Ser 115 120 125 att tgggac aac ctt aca tgg cag caa tgg gac caa cac ata aac aat 431 Ile Trp AspAsn Leu Thr Trp Gln Gln Trp Asp Gln His Ile Asn Asn 130 135 140 gta agctcc att ata tat gat gaa ata caa gca gca caa gac caa cag 479 Val Ser SerIle Ile Tyr Asp Glu Ile Gln Ala Ala Gln Asp Gln Gln 145 150 155 gaa aagaat gta aaa gca ttg ttg gag cta gat gaa tgg gcc tct ctt 527 Glu Lys AsnVal Lys Ala Leu Leu Glu Leu Asp Glu Trp Ala Ser Leu 160 165 170 175 tggaat tgg ttt gac ata act aaa tgg ttg tgg tat ata aaa ata gct 575 Trp AsnTrp Phe Asp Ile Thr Lys Trp Leu Trp Tyr Ile Lys Ile Ala 180 185 190 ataatc ata gtg gga gca cta ata ggt ata aga gtt atc atg ata gta 623 Ile IleIle Val Gly Ala Leu Ile Gly Ile Arg Val Ile Met Ile Val 195 200 205 cttaat cta gtg aag aac att agg cag gga tat caa ccc ctc tcg ttg 671 Leu AsnLeu Val Lys Asn Ile Arg Gln Gly Tyr Gln Pro Leu Ser Leu 210 215 220 cagatc cct gtc cca cac cgg cag gaa gca gaa acg cca gga aga aca 719 Gln IlePro Val Pro His Arg Gln Glu Ala Glu Thr Pro Gly Arg Thr 225 230 235 ggagaa gaa ggt gga gaa gga gac agg ccc aag tgg aca gcc ttg cca 767 Gly GluGlu Gly Gly Glu Gly Asp Arg Pro Lys Trp Thr Ala Leu Pro 240 245 250 255cca gga ttc ttg caa cag ttg tac acg gat ctc agg aca ata atc ttg 815 ProGly Phe Leu Gln Gln Leu Tyr Thr Asp Leu Arg Thr Ile Ile Leu 260 265 270tgg act tac cac ctc ttg agc aac tta ata tca ggg atc cgg agg ctg 863 TrpThr Tyr His Leu Leu Ser Asn Leu Ile Ser Gly Ile Arg Arg Leu 275 280 285atc gac tac ctg gga ctg gga ctg tgg atc ctg gga caa aag aca att 911 IleAsp Tyr Leu Gly Leu Gly Leu Trp Ile Leu Gly Gln Lys Thr Ile 290 295 300gaa gct tgt aga ctt tgt gga gct gta atg caa tat tgg cta caa gaa 959 GluAla Cys Arg Leu Cys Gly Ala Val Met Gln Tyr Trp Leu Gln Glu 305 310 315ttg aaa aat agt gct aca aac ctg ctt gat act att gca gtg tca gtt 1007 LeuLys Asn Ser Ala Thr Asn Leu Leu Asp Thr Ile Ala Val Ser Val 320 325 330335 gcc aat tgg act gac ggc atc atc tta ggt cta caa aga ata gga caa 1055Ala Asn Trp Thr Asp Gly Ile Ile Leu Gly Leu Gln Arg Ile Gly Gln 340 345350 gg 1057 45 1057 DNA Human immunodeficiency virus 45 ccttgtcctattctttgtag acctaagatg atgccgtcag tccaattggc aactgcaact 60 gcaatagtatcaagcaggtt tgtagcacta tttttcaatt cttgtagcca atattgcatt 120 acagctccacaaagtctaca agcttcaatt gtcttttgtc ccaggatcca cagtcccagt 180 cccaggtagtcgatcagcct ccggatccct gatattaagt tgctcaagag gtggtaagtc 240 cacaagattattgtcctgag atccgtgtac aactgttgca agaatcctgg tggcaaggct 300 gtccacttgggcctgtctcc ttctccacct tcttctcctg ttcttcctgg cgtttctgct 360 tcctgccggtgtgggacagg gatctgcaac gagaggggtt gatatccctg cctaatgttc 420 ttcactagattaagtactat catgataact cttataccta ttagtgctcc cactatgatt 480 atagctatttttatatacca caaccattta gttatgtcaa accaattcca aagagaggcc 540 cattcatctagctccaacaa tgcttttaca ttcttttcct gttggtcttg tgctgcttgt 600 atttcatcatatataatgga gcttacattg tttatgtgtt ggtcccattg ctgccatgta 660 aggttgtcccaaatactgtc atcattatat cctcctgacc atgatgtgtt ccattttact 720 gatgtgtaacagattagttt tcctttacag ccccataggt ttaggcgttg ctgattctgt 780 ataagggtttctaaggcttg caggcgagct cggagttgtc taatacccca tacagataac 840 ctcagcaagtgttgctgggc ctgtatcgct ctcagcaggt tgtcctgctg ttgcactata 900 cccttcagtacactgtgggt ccgtaccgtc agcgctgttg ccgctgcgcc catagtgcta 960 cctgctgcacttagcacccc caagaatagc attcccaatc ctactgctct tttttccctg 1020 tgaggggtgtgaatgtttat tattggtctt gacattt 1057 46 351 PRT Human immunodeficiencyvirus 46 Met Ser Arg Pro Ile Ile Asn Ile His Thr Pro His Arg Glu Lys Arg1 5 10 15 Arg Val Gly Leu Gly Met Leu Phe Leu Gly Val Leu Ser Ala AlaGly 20 25 30 Ser Thr Met Gly Ala Ala Ala Thr Ala Leu Thr Val Arg Thr HisSer 35 40 45 Val Leu Lys Gly Ile Val Gln Gln Gln Asp Asn Leu Leu Arg AlaIle 50 55 60 Gln Ala Gln Gln His Leu Leu Arg Leu Ser Val Trp Gly Ile ArgGln 65 70 75 80 Leu Arg Ala Arg Leu Gln Ala Leu Glu Thr Leu Ile Gln AsnGln Gln 85 90 95 Arg Leu Asn Leu Trp Gly Cys Lys Gly Lys Leu Ile Cys TyrThr Ser 100 105 110 Val Lys Trp Asn Thr Ser Trp Ser Gly Gly Tyr Asn AspAsp Ser Ile 115 120 125 Trp Asp Asn Leu Thr Trp Gln Gln Trp Asp Gln HisIle Asn Asn Val 130 135 140 Ser Ser Ile Ile Tyr Asp Glu Ile Gln Ala AlaGln Asp Gln Gln Glu 145 150 155 160 Lys Asn Val Lys Ala Leu Leu Glu LeuAsp Glu Trp Ala Ser Leu Trp 165 170 175 Asn Trp Phe Asp Ile Thr Lys TrpLeu Trp Tyr Ile Lys Ile Ala Ile 180 185 190 Ile Ile Val Gly Ala Leu IleGly Ile Arg Val Ile Met Ile Val Leu 195 200 205 Asn Leu Val Lys Asn IleArg Gln Gly Tyr Gln Pro Leu Ser Leu Gln 210 215 220 Ile Pro Val Pro HisArg Gln Glu Ala Glu Thr Pro Gly Arg Thr Gly 225 230 235 240 Glu Glu GlyGly Glu Gly Asp Arg Pro Lys Trp Thr Ala Leu Pro Pro 245 250 255 Gly PheLeu Gln Gln Leu Tyr Thr Asp Leu Arg Thr Ile Ile Leu Trp 260 265 270 ThrTyr His Leu Leu Ser Asn Leu Ile Ser Gly Ile Arg Arg Leu Ile 275 280 285Asp Tyr Leu Gly Leu Gly Leu Trp Ile Leu Gly Gln Lys Thr Ile Glu 290 295300 Ala Cys Arg Leu Cys Gly Ala Val Met Gln Tyr Trp Leu Gln Glu Leu 305310 315 320 Lys Asn Ser Ala Thr Asn Leu Leu Asp Thr Ile Ala Val Ser ValAla 325 330 335 Asn Trp Thr Asp Gly Ile Ile Leu Gly Leu Gln Arg Ile GlyGln 340 345 350 47 22 DNA Artificial Sequence Description of ArtificialSequence primer 47 ctagcagtgg cgcccgaaca gg 22 48 23 DNA ArtificialSequence Description of Artificial Sequence primer 48 aatgaggaagcwgcagawtg gga 23 49 23 DNA Artificial Sequence Description ofArtificial Sequence primer 49 tcccawtctg cwgcttcctc att 23 50 26 DNAArtificial Sequence Description of Artificial Sequence primer 50ccaaggggaa gtgacatagc aggaac 26 51 21 DNA Artificial SequenceDescription of Artificial Sequence primer 51 cgttgttcag aattcaaacc c 2152 21 DNA Artificial Sequence Description of Artificial Sequence primer52 tccctaaaaa attagcctgt c 21 53 19 DNA Artificial Sequence Descriptionof Artificial Sequence primer 53 aaacctccaa ttcccccta 19 54 39 PRT Humanimmunodeficiency virus 54 Cys Ile Arg Glu Gly Ile Ala Glu Val Gln AspIle Tyr Thr Gly Pro 1 5 10 15 Met Arg Trp Arg Ser Met Thr Leu Lys ArgSer Asn Asn Thr Ser Pro 20 25 30 Arg Ser Arg Val Ala Tyr Cys 35 55 41PRT Human immunodeficiency virus 55 Cys Ile Arg Glu Gly Ile Ala Glu ValGln Asp Leu His Thr Gly Pro 1 5 10 15 Leu Arg Trp Arg Ser Met Thr LeuLys Lys Ser Ser Asn Ser His Thr 20 25 30 Gln Pro Arg Ser Lys Val Ala TyrCys 35 40 56 9793 DNA Human immunodeficiency virus 56 ctggatgggttaatttactc ccataagaga gcagaaatcc tggatctctg gatatatcac 60 actcagggattcttccctga ttggcagtgt tacacaccgg gaccaggacc tagattccca 120 ctgacatttggatggttgtt taaactggta ccagtgtcag cagaagaggc agagagactg 180 ggtaatacaaatgaagatgc tagtcttcta catccagctt gtaatcatgg agctgaggat 240 gcacacggggagatactaaa atggcagttt gatagatcat taggcttaac acatatagcc 300 ctgcaaaagcacccagagct cttccccaag taactgacac tgcgggactt tccagactgc 360 tgacactgcggggactttcc agcgtgggag ggataagggg cggttcgggg agtggctaac 420 cctcagatgctgcatataag cagctgcttt ccgcttgtac cgggtcttag ttagaggacc 480 aggtctgagcccgggagctc cctggcctct agctgaaccc gctgcttaac gctcaataaa 540 gcttgccttgagtgagaagc agtgtgtgct catctgttca accctggtgt ctagagatcc 600 ctcagatcacttagactgaa gcagaaaatc tctagcagtg gcgcccgaac agggacgcga 660 aagtgaaagtggaaccaggg aagaaaacct ccgacgcaac gggctcggct tagcggagtg 720 cacctgctaagaggcgagag gaactcacaa gagggtgagt aaatttgctg gcggtggcca 780 gacctaggggaagggcgaag tccctagggg aggaagatgg gtgcgagagc gtctgtgttg 840 acagggagtaaattggatgc atgggaacga attaggttaa ggccaggatc taaaaaggca 900 tataggctaaaacatttagt atgggcaagc agggagctgg aaagatacgc atgtaatcct 960 ggtctattagaaactgcaga aggtactgag caactgctac agcagttaga gccagctctc 1020 aagacagggtcagaggacct gaaatctctc tggaacgcaa tagcagtact ctggtgcgtt 1080 cacaacagatttgacatccg agatacacag caggcaatac aaaagttaaa ggaagtaatg 1140 gcaagcaggaagtctgcaga ggccgctaag gaagaaacaa gccctaggca gacaagtcaa 1200 aattaccctatagtaacaaa tgcacaggga caaatggtac atcaagccat ctcccccagg 1260 actttaaatgcatgggtaaa ggcagtagaa gagaaggcct ttaaccctga aattattcct 1320 atgtttatggcattatcaga aggggctgtc ccctatgata tcaataccat gctgaatgcc 1380 atagggggacaccaaggggc tttacaagtg ttgaaggaag taatcaatga ggaagcagca 1440 gaatgggatagaactcatcc accagcaatg gggccgttac caccagggca gataagggaa 1500 ccaacaggaagtgacattgc tggaacaact agcacacagc aagagcaaat tatatggact 1560 actagaggggctaactctat cccagtagga gacatctata gaaaatggat agtgctagga 1620 ctaaacaaaatggtaaaaat gtacagtcca gtgagcatct tagatattag gcagggacca 1680 aaagaaccattcagagatta tgtagatcgg ttttacaaaa cattaagagc tgagcaagct 1740 actcaagaagtaaagaattg gatgacagaa accttgcttg ttcagaattc aaacccagat 1800 tgtaaacaaattctgaaagc attaggacca gaagctactt tagaagaaat gatggtagcc 1860 tgtcaaggagtaggagggcc aactcacaag gcaaaaatac tagcagaagc aatggcttct 1920 gcccagcaagatttaaaagg aggatacaca gcagtattca tgcaaagagg gcagaatcca 1980 aatagaaaagggcccataaa atgcttcaat tgtggaaaag agggacatat agcaaaaaac 2040 tgtcgagcacctagaaaaag gggttgctgg aaatgtggac aggaaggtca ccaaatgaaa 2100 gattgcaaaaatggaagaca ggcaaatttt ttagggaagt actggcctcc ggggggcacg 2160 aggccaggcaattatgtgca gaaacaagtg tccccatcag ccccaccaat ggaggaggca 2220 gtgaaggaacaagagaatca gagtcagaag ggggatcagg aagagctgta cccatttgcc 2280 tccctcaaatccctctttgg gacagaccaa tagtcacagc aaaggttggg ggtcatctat 2340 gtgaggctttactggataca ggggcagatg atacagtatt aaataacata caattagaag 2400 gaagatggacaccaaaaatg atagggggta taggaggctt tataaaagta aaagagtata 2460 acaatgtgacagtagaagta caaggaaagg aagtacaggg aacagtattg gtgggaccta 2520 ctcctgttaatattcttggg agaaacatat tgacaggatt aggatgtaca ctaaatttcc 2580 ctataagtcccatagcccca gtgccagtaa agctaaaacc aggaatggat ggaccaaaag 2640 taaaacaatggcccctatct agagagaaaa tagaagcact aactgcaata tgtcaagaaa 2700 tggaacaggaaggaaaaatc tcaagaatag gacctgaaaa tccttataat acacctattt 2760 ttgctataaaaaagaaagat agcactaagt ggagaaaatt ggtagacttc agagaattaa 2820 ataaaagaacacaagatttc tgggaggtgc aattaggtat tccacatcca gggggtttaa 2880 agcaaaggcaatctgttaca gtcttagatg taggagatgc ttatttctca tgccctttag 2940 atccagactttagaaaatac actgccttca ctattcctag tgtgaacaat gagaccccag 3000 gagtaagataccagtacaat gtcctcccgc aagggtggaa aggttcacca gccatatttc 3060 agagttcaatgacaaagatt ctagatccat ttagaaaaag caacccagaa gtagaaattt 3120 atcagtacatagatgactta tatgtaggat cagatttacc attggcagaa catagaaaga 3180 gggtcgaattgcttagggaa catttatatc agtggggatt tactacccct gataaaaagc 3240 atcagaaggaacctcccttt ttatggatgg gatatgagct ccacccagac aagtggacag 3300 tacagcccatccaattgcct gacaaagaag tgtggacagt aaatgatata caaaaattag 3360 taggaaaattaaattgggca agtcaaatct atcaaggaat tagagtaaaa gaattgtgca 3420 agttaatcagaggaaccaaa tcattgacag aggtagtacc tttaagtaaa gaggcagaac 3480 tagaattagaagaaaacaga gaaaagctaa aagagccagt acatggagta tattaccagc 3540 ctgacaaagacttgtgggtt agtattcaga agcatggaga agggcaatgg acttaccagg 3600 tatatcaggatgaacataag aaccttaaaa caggaaaata tgctaggcaa aaggcctccc 3660 acacaaatgatataagacaa ttggcagaag tagtccagaa ggtgtctcaa gaagctatag 3720 ttatatgggggaaattacct aaattcaggc tgccagttac tagagaaact tgggaaactt 3780 ggtgggcagaatattggcag gccacctgga ttcctgaatg ggaatttgtc agcacacccc 3840 cattgatcaaattatggtac cagttagaaa cagaacctat tgtaggggca gaaacctttt 3900 atgtagatggagcagctaat aggaatacaa aactaggaaa ggcgggatat gttacagaac 3960 aaggaaaacagaacataata aagttagaag agacaaccaa tcaaaaggct gaattaatgg 4020 ctgtattaatagccttgcag gattccaagg agcaagtaaa catagtaaca gactcacaat 4080 atgtattgggcatcatatcc tcccaaccaa cacagagtga ctcccctata gttcagcaga 4140 taatagaggaactaacaaaa aaggaacgag tgtatcttac atgggttcct gctcacaaag 4200 gcataggaggaaatgaaaaa atagataaat tagtaagcaa agacattaga agagtcctgt 4260 tcctggaaggaatagatcag gcacaagaag atcatgaaaa atatcatagt aattggagag 4320 cattagctagtgactttgga ttaccaccaa tagtagccaa ggaaatcatt gctagttgtc 4380 ctaaatgccatataaaaggg gaagcaacgc atggtcaagt agactacagc ccagagatat 4440 ggcaaatggattgtacacat ttagaaggca aaatcataat agttgctgtc catgtagcaa 4500 gtgactttatagaagcagag gtgataccag cagaaacagg acaggaaact gcctatttcc 4560 tgttaaaattagcagcaaga tggcctgtca aagtaataca tacagacaat ggacctaatt 4620 ttacaagtgcagccatgaaa gctgcatgtt ggtggacagg catacaacat gagtttggga 4680 taccatataatccacaaagt caaggagtag tagaagccat gaataaagaa ttaaaatcta 4740 ttatacagcaggtgagggac caagcagagc atttaaaaac agcagtacaa atggcagtct 4800 ttgttcacaattttaaaaga aaagggggga ttggggggta cactgcaggg gagagactaa 4860 tagacatactagcatcacaa atacaaacaa cagaactaca aaaacaaatt ttaaaaatca 4920 acaattttcgggtctattac agagatagca gagaccctat ttggaaagga ccggcacaac 4980 tcctgtggaaaggtgagggg gcagtagtca tacaagataa aggagacatt aaagtggtac 5040 caagaagaaaggcaaaaata atcagagatt atggaaaaca gatggcaggt actgatagta 5100 tggcaaatagacagacagaa agtgaaagca tggaacagcc tggtgaaata ccataaatac 5160 atgtctaagaaggccgcgaa ctggcgttat aggcatcatt atgaatccag gaatccaaaa 5220 gtcagttcggcggtgtatat tccagtagca gaagctgata tagtggtcac cacatattgg 5280 ggattaatgccaggggaaag agaggaacac ttgggacatg gggttagtat agaatggcaa 5340 tacaaggagtataaaacaca gattgatcct gaaacagcag acaggatgat acatctgcat 5400 tatttcacatgttttacaga atcagcaatc aggaaggcca ttctagggca gagagtgctg 5460 accaagtgtgaatacctggc aggacatagt caggtaggga cactacaatt cttagccttg 5520 aaagcagtagtgaaagtaaa aagaaataag cctcccctac ccagtgtcca gagattaaca 5580 gaagatagatggaacaagcc ctggaaaatc agggaccagc tagggagcca ttcaatgaat 5640 ggacactagagctcctggaa gagctgaaag aagaagcagt aagacatttc cctaggcctt 5700 ggttacaagcctgtgggcag tacatttatg agacttatgg agacacttgg gaaggagtta 5760 tggcaattataagaatctta caacaactac tgtttaccca ttatagaatt ggatgccaac 5820 atagtagaataggaattctc ccatctaaca caagaggaag aggaagaaga aatggatcca 5880 gtagatcctgagatgccccc ttggcatcac cctgggagca agccccaaac cccttgtaat 5940 aattgctattgcaaaagatg ctgctatcat tgctatgttt gtttcacaaa gaagggtttg 6000 ggaatctcccatggcaggaa gaagcgaaga agaccagcag ctgctgcaag ctatccagat 6060 aataaagatcctgtaccaga gcagtaagta acgctgatgc atcaagagaa cctgctagcc 6120 ttaatagctttaagtgcttt gtgtcttata aatgtactta tatggttgtt taaccttaga 6180 atttatttagtgcaaagaaa acaagataga agggagcagg aaatacttga aagattaagg 6240 agaataaaggaaatcaggga tgacagtgac tatgaaagta atgaagaaga acaacaggaa 6300 gtcatggagcttatacatag ccatggcttt gctaatccca tgtttgagtt atagtaaaca 6360 attgtatgccacagtttatt ctggggtacc tgtatgggaa gaggcagcac cagtactatt 6420 ctgtgcttcagatgctaacc taacaagcac tgaacagcat aatatttggg catcacaagc 6480 ctgcgttcctacagatccca atccacatga atttccacta ggcaatgtga cagataactt 6540 tgatatatggaaaaattaca tggtggacca aatgcatgaa gacatcatta gtttgtggga 6600 acagagtttaaagccttgtg agaaaatgac tttcttatgt gtacaaatga actgtgtaga 6660 tctgcaaacaaataaaacag gcctattaaa tgagacaata aatgagatga gaaattgtag 6720 ttttaatgtaactacagtcc tcacagacaa aaaggagcaa aaacaggctc tattctatgt 6780 atcagatctgagtaaggtta atgactcaaa tgcagtaaat ggaacaacat atatgttaac 6840 taattgtaactccacaatta tcaagcaggc ctgtcccaag gtaagttttg agcccattcc 6900 catacactattgtgctccaa caggatatgc catctttaag tgtaatgaca cagactttaa 6960 tggaacaggcctatgccaca atatttcagt ggttacttgt acacatggca tcaagccaac 7020 agtaagtactcaactaatac tgaatgggac actctctaga gaaaagataa gaattatggg 7080 aaaaaatattacagaatcag caaagaatat catagtaacc ctaaacactc ctataaacat 7140 gacctgcataagagaaggaa ttgcagaggt acaagatata tatacaggtc caatgagatg 7200 gcgcagtatgacacttaaaa gaagtaacaa tacatcacca agatcaaggg tagcttattg 7260 tacatataataagactgtat gggaaaatgc cctacaacaa acagctataa ggtatttaaa 7320 tcttgtaaaccaaacagaga atgttaccat aatattcagc agaactagtg gtggagatgc 7380 agaagtaagccatttacatt ttaactgtca tggagaattc ttttattgta acacatctgg 7440 gatgtttaactatactttta tcaactgtac aaagtccgga tgccaggaga tcaaagggag 7500 caatgagaccaataaaaatg gtactatacc ttgcaagtta agacagctag taagatcatg 7560 gatgaagggagagtcgagaa tctatgcacc tcccatcccc ggcaacttaa catgtcattc 7620 caacataactggaatgattc tacagttaga tcaaccatgg aattccacag gtgaaaatac 7680 acttagaccagtagggggag atatgaaaga tatatggaga actaaattgt acaactacaa 7740 agtagtacagataaaacctt ttagtgtagc acctacaaaa atgtcaagac caataataaa 7800 cattcacacccctcacaggg aaaaaagagc agtaggattg ggaatgctat tcttgggggt 7860 gctaagtgcagcaggtagca ctatgggcgc agcggcaaca gcgctgacgg tacggaccca 7920 cagtgtactgaagggtatag tgcaacagca ggacaacctg ctgagagcga tacaggccca 7980 gcaacacttgctgaggttat ctgtatgggg tattagacaa ctccgagctc gcctgcaagc 8040 cttagaaacccttatacaga atcagcaacg cctaaaccta tggggctgta aaggaaaact 8100 aatctgttacacatcagtaa aatggaacac atcatggtca ggaagatata atgatgacag 8160 tatttgggacaaccttacat ggcagcaatg ggaccaacac ataaacaatg taagctccat 8220 tatatatgatgaaatacaag cagcacaaga ccaacaggaa aagaatgtaa aagcattgtt 8280 ggagctagatgaatgggcct ctctttggaa ttggtttgac ataactaaat ggttgtggta 8340 tataaaaatagctataatca tagtgggagc actaataggt ataagagtta ttatgataat 8400 acttaatctagtgaagaaca ttaggcaggg atatcaaccc ctctcgttgc agatccctgt 8460 cccacaccggcaggaagcag aaacgccagg aagaacagga gaagaaggtg gagaaggaga 8520 caggcccaagtggacagcct tgccaccagg attcttgcaa cagttgtaca cggatctcag 8580 gacaataatcttgtggactt accacctctt gagcaactta atatcaggga tccggaggct 8640 gatcgactacctgggactgg gactgtggat cctgggacaa aagacaattg aagcttgtag 8700 actttgtggagctgtaatgc aatattggct acaagaattg aaaaatagtg ctacaaacct 8760 gcttgatactattgcagtgt cagttgccaa ttggactgac ggcatcatct taggtctaca 8820 aagaataggacaaggattcc ttcacatccc aagaagaatt agacaaggtg cagaaagaat 8880 cttagtgtaacatggggaat gcatggagca aaagcaaatt tgcaggatgg tcagaagtaa 8940 gagatagaatgagacgatcc tcctctgatc ctcaacaacc atgtgcacct ggagtaggag 9000 ctgtctccagggagttagca actagagggg gaatatcaag ttcccacact cctcaaaaca 9060 atgcagcccttgcattccta gacagccaca aagatgagga tgtaggcttc ccagtaagac 9120 ctcaagtgcctctaaggcca atgaccttta aagcagcctt tgacctcagc ttctttttaa 9180 aagaaaagggaggactggat gggttaattt actcccataa gagagcagaa atcctggatc 9240 tctggatatatcacactcag ggattcttcc ctgattggca gtgttacaca ccgggaccag 9300 gacctagattcccactgaca tttggatggt tgtttaaact ggtaccagtg tcagcagaag 9360 aggcagagagactgggtaat acaaatgaag atgctagtct tctacatcca gcttgtaatc 9420 atggagctgaggatgcacac ggggagatac taaaatggca gtttgataga tcattaggct 9480 taacacatatagccctgcaa aagcacccag agctcttccc caagtaactg acactgcggg 9540 actttccagactgctgacac tgcggggact ttccagcgtg ggagggataa ggggcggttc 9600 ggggagtggctaaccctcag atgctgcata taagcagctg ctttccgctt gtaccgggtc 9660 ttagttagaggaccaggtct gagcccggga gctccctggc ctctagctga acccgctgct 9720 taacgctcaataaagcttgc cttgagtgag aagcagtgtg tgctcatctg ttcaaccctg 9780 gtgtctagagatc 9793 57 1733 DNA Human immunodeficiency virus 57 aaacctccgacgcaacgggc tcggcttagc ggagtgcacc tgctaagagg cgagaggaac 60 tcacaagagggtgagtaaat ttgctggcgg tggccagacc taggggaagg gcgaagtccc 120 taggggaggaagatgggtgc gagagcgtct gtgttgacag ggagtaaatt ggatgcatgg 180 gaacgaattaggttaaggcc aggatctaaa aaggcatata ggctaaaaca tttagtatgg 240 gcaagcagggagctggaaag atacgcatgt aatcctggtc tattagaaac tgcagaaggt 300 actgagcaactgctacagca gttagagcca gctctcaaga cagggtcaga ggacctgaaa 360 tctctctggaacgcaatagc agtactctgg tgcgttcaca acagatttga catccgagat 420 acacagcaggcaatacaaaa gttaaaggaa gtaatggcaa gcaggaagtc tgcagaggcc 480 gctaaggaagaaacaagccc taggcagaca agtcaaaatt accctatagt aacaaatgca 540 cagggacaaatggtacatca agccatctcc cccaggactt taaatgcatg ggtaaaggca 600 gtagaagagaaggcctttaa ccctgaaatt attcctatgt ttatggcatt atcagaaggg 660 gctgtcccctatgatatcaa taccatgctg aatgccatag ggggacacca aggggcttta 720 caagtgttgaaggaagtaat caatgaggaa gcagcagaat gggatagaac tcatccacca 780 gcaatggggccgttaccacc agggcagata agggaaccaa caggaagtga cattgctgga 840 acaactagcacacagcaaga gcaaattata tggactacta gaggggctaa ctctatccca 900 gtaggagacatctatagaaa atggatagtg ctaggactaa acaaaatggt aaaaatgtac 960 agtccagtgagcatcttaga tattaggcag ggaccaaaag aaccattcag agattatgta 1020 gatcggttttacaaaacatt aagagctgag caagctactc aagaagtaaa gaattggatg 1080 acagaaaccttgcttgttca gaattcaaac ccagattgta aacaaattct gaaagcatta 1140 ggaccagaagctactttaga agaaatgatg gtagcctgtc aaggagtagg agggccaact 1200 cacaaggcaaaaatactagc agaagcaatg gcttctgccc agcaagattt aaaaggagga 1260 tacacagcagtattcatgca aagagggcag aatccaaata gaaaagggcc cataaaatgc 1320 ttcaattgtggaaaagaggg acatatagca aaaaactgtc gagcacctag aaaaaggggt 1380 tgctggaaatgtggacagga aggtcaccaa atgaaagatt gcaaaaatgg aagacaggca 1440 aattttttagggaagtactg gcctccgggg ggcacgaggc caggcaatta tgtgcagaaa 1500 caagtgtccccatcagcccc accaatggag gaggcagtga aggaacaaga gaatcagagt 1560 cagaagggggatcaggaaga gctgtaccca tttgcctccc tcaaatccct ctttgggaca 1620 gaccaatagtcacagcaaag gttgggggtc atctatgtga ggctttactg gatacagggg 1680 cagatgatacagtattaaat aacatacaat tagaaggaag atggacacca aaa 1733 58 1733 DNA Humanimmunodeficiency virus 58 aaacctccaa cgcaacgggc tcggcttagc ggagtgcacctgctaagagg cgagaggaac 60 tcacaagagg gtgagtaaat ttgctggcgg tggccagacctaggggaagg gcgaagtccc 120 taggggagga agatgggtgc gagacggtct gtgttgacagggagtaaatt ggatgcatgg 180 gaacgaatta ggttaaggcc aggatctaaa aaggcatataggctaaaaca tttagtatgg 240 gcaagcaggg agctggaaag atacgcatat aatcctggtctactagaaac tgcagaaggt 300 actgaacaac tgctacagca gttagagcca gctctcaagacagggtcaga ggacctgaaa 360 tccctctgga acgcaatagc agtactctgg tgcgttcacaacagatttga catccgagat 420 acacagcagg caatacaaaa gttaaaggaa gtaatggcaagcaggaagtc tgcagaggcc 480 gctaaggaag aaacaagctc aaggcaggca agtcaaaattaccctatagt aacaaatgca 540 cagggacaaa tggtacatca agccatatcc cctaggactttaaatgcatg ggtaaaggca 600 gtagaagaaa aggcctttaa ccctgaaatt attcctatgtttatggcatt atcagaaggg 660 gctgtcccct atgatatcaa taccatgctg aatgccatagggggacacca aggggcttta 720 caagtgttga aggaagtaat caatgaggaa gcagcagattgggatagaac tcatccacca 780 gcaatggggc cgttaccacc agggcagata agggaaccaacaggaagtga cattgctgga 840 acaactagca cacagcaaga gcaaattata tggactactagaggggctaa ctctatccca 900 gtaggagaca tctatagaaa atggatagtg ttaggactaaacaaaatggt aaaaatgtac 960 agtccagtga gcatcttaga tattaggcag ggaccaaaagaaccattcag agattatgta 1020 gatcggtttt acaaaacatt aagagctgag caagctactcaagaagtaaa gaattggatg 1080 acagaaaccc tcgttgttca gaattcaaac ccagattgtaaacaaattct gaaagcatta 1140 ggaccaggag ctactttaga agaaatgatg gtagcctgtcaaggagtagg agggccaact 1200 cacaaggcaa aaatactagc agaagcaatg gcttctgcccagcaagattt aaagggagga 1260 tacacagcag tattcatgca aagagggcag aatccaaatagaaaagggcc tataaaatgt 1320 ttcaattgtg gaaaagaggg acatatagca aaaaactgtcgagcacctag aagaaggggt 1380 tactggaaat gtggacagga aggtcaccaa atgaaagattgcaaaaatgg aagacaggct 1440 atttttttag ggaagtactg gcctccgggg ggcacgaggccagccaatta tgtgcagaaa 1500 caagtgtccc catcagcccc accaatggag gaggcagtgaaggaacaaga gaatcagaat 1560 caaaaggggg atcaggaaga gctgtaccca tttgcctccctcaaatccct ctttgggaca 1620 gaccaatagt cacagcaaag gttgggggcc atctatgtgaggctttactg gatacagggg 1680 cagatgatac agtattaaat aacatacaat tagaaggaagatggacaccc aaa 1733 59 498 PRT Human immunodeficiency virus 59 Met GlyAla Arg Ala Ser Val Leu Thr Gly Ser Lys Leu Asp Ala Trp 1 5 10 15 GluArg Ile Arg Leu Arg Pro Gly Ser Lys Lys Ala Tyr Arg Leu Lys 20 25 30 HisLeu Val Trp Ala Ser Arg Glu Leu Glu Arg Tyr Ala Cys Asn Pro 35 40 45 GlyLeu Leu Glu Thr Ala Glu Gly Thr Glu Gln Leu Leu Gln Gln Leu 50 55 60 GluPro Ala Leu Lys Thr Gly Ser Glu Asp Leu Lys Ser Leu Trp Asn 65 70 75 80Ala Ile Ala Val Leu Trp Cys Val His Asn Arg Phe Asp Ile Arg Asp 85 90 95Thr Gln Gln Ala Ile Gln Lys Leu Lys Glu Val Met Ala Ser Arg Lys 100 105110 Ser Ala Glu Ala Ala Lys Glu Glu Thr Ser Pro Arg Gln Thr Ser Gln 115120 125 Asn Tyr Pro Ile Val Thr Asn Ala Gln Gly Gln Met Val His Gln Ala130 135 140 Ile Ser Pro Arg Thr Leu Asn Ala Trp Val Lys Ala Val Glu GluLys 145 150 155 160 Ala Phe Asn Pro Glu Ile Ile Pro Met Phe Met Ala LeuSer Glu Gly 165 170 175 Ala Val Pro Tyr Asp Ile Asn Thr Met Leu Asn AlaIle Gly Gly His 180 185 190 Gln Gly Ala Leu Gln Val Leu Lys Glu Val IleAsn Glu Glu Ala Ala 195 200 205 Glu Trp Asp Arg Thr His Pro Pro Ala MetGly Pro Leu Pro Pro Gly 210 215 220 Gln Ile Arg Glu Pro Thr Gly Ser AspIle Ala Gly Thr Thr Ser Thr 225 230 235 240 Gln Gln Glu Gln Ile Ile TrpThr Thr Arg Gly Ala Asn Ser Ile Pro 245 250 255 Val Gly Asp Ile Tyr ArgLys Trp Ile Val Leu Gly Leu Asn Lys Met 260 265 270 Val Lys Met Tyr SerPro Val Ser Ile Leu Asp Ile Arg Gln Gly Pro 275 280 285 Lys Glu Pro PheArg Asp Tyr Val Asp Arg Phe Tyr Lys Thr Leu Arg 290 295 300 Ala Glu GlnAla Thr Gln Glu Val Lys Asn Trp Met Thr Glu Thr Leu 305 310 315 320 LeuVal Gln Asn Ser Asn Pro Asp Cys Lys Gln Ile Leu Lys Ala Leu 325 330 335Gly Pro Glu Ala Thr Leu Glu Glu Met Met Val Ala Cys Gln Gly Val 340 345350 Gly Gly Pro Thr His Lys Ala Lys Ile Leu Ala Glu Ala Met Ala Ser 355360 365 Ala Gln Gln Asp Leu Lys Gly Gly Tyr Thr Ala Val Phe Met Gln Arg370 375 380 Gly Gln Asn Pro Asn Arg Lys Gly Pro Ile Lys Cys Phe Asn CysGly 385 390 395 400 Lys Glu Gly His Ile Ala Lys Asn Cys Arg Ala Pro ArgLys Arg Gly 405 410 415 Cys Trp Lys Cys Gly Gln Glu Gly His Gln Met LysAsp Cys Lys Asn 420 425 430 Gly Arg Gln Ala Asn Phe Leu Gly Lys Tyr TrpPro Pro Gly Gly Thr 435 440 445 Arg Pro Gly Asn Tyr Val Gln Lys Gln ValSer Pro Ser Ala Pro Pro 450 455 460 Met Glu Glu Ala Val Lys Glu Gln GluAsn Gln Ser Gln Lys Gly Asp 465 470 475 480 Gln Glu Glu Leu Tyr Pro PheAla Ser Leu Lys Ser Leu Phe Gly Thr 485 490 495 Asp Gln 60 498 PRT Humanimmunodeficiency virus 60 Met Gly Ala Arg Arg Ser Val Leu Thr Gly SerLys Leu Asp Ala Trp 1 5 10 15 Glu Arg Ile Arg Leu Arg Pro Gly Ser LysLys Ala Tyr Arg Leu Lys 20 25 30 His Leu Val Trp Ala Ser Arg Glu Leu GluArg Tyr Ala Tyr Asn Pro 35 40 45 Gly Leu Leu Glu Thr Ala Glu Gly Thr GluGln Leu Leu Gln Gln Leu 50 55 60 Glu Pro Ala Leu Lys Thr Gly Ser Glu AspLeu Lys Ser Leu Trp Asn 65 70 75 80 Ala Ile Ala Val Leu Trp Cys Val HisAsn Arg Phe Asp Ile Arg Asp 85 90 95 Thr Gln Gln Ala Ile Gln Lys Leu LysGlu Val Met Ala Ser Arg Lys 100 105 110 Ser Ala Glu Ala Ala Lys Glu GluThr Ser Ser Thr Gln Ala Ser Gln 115 120 125 Asn Tyr Pro Ile Val Thr AsnAla Gln Gly Gln Met Val His Gln Ala 130 135 140 Ile Ser Pro Arg Thr LeuAsn Ala Trp Val Lys Ala Val Glu Glu Lys 145 150 155 160 Ala Phe Asn ProGlu Ile Ile Pro Met Phe Met Ala Leu Ser Glu Gly 165 170 175 Ala Val ProTyr Asp Ile Asn Thr Met Leu Asn Ala Ile Gly Gly His 180 185 190 Gln GlyAla Leu Gln Val Leu Lys Glu Val Ile Asn Glu Glu Ala Ala 195 200 205 AspTrp Asp Arg Thr His Pro Pro Ala Met Gly Pro Leu Pro Pro Gly 210 215 220Gln Ile Arg Glu Pro Thr Gly Ser Asp Ile Ala Gly Thr Thr Ser Thr 225 230235 240 Gln Gln Glu Gln Ile Ile Trp Thr Thr Arg Gly Ala Asn Ser Ile Pro245 250 255 Val Gly Asp Ile Tyr Arg Lys Trp Ile Val Leu Gly Leu Asn LysMet 260 265 270 Val Lys Met Tyr Ser Pro Val Ser Ile Leu Asp Ile Arg GlnGly Pro 275 280 285 Lys Glu Pro Phe Arg Asp Tyr Val Asp Arg Phe Tyr LysThr Leu Arg 290 295 300 Ala Glu Gln Ala Thr Gln Glu Val Lys Asn Trp MetThr Glu Thr Leu 305 310 315 320 Val Val Gln Asn Ser Asn Pro Asp Cys LysGln Ile Leu Lys Ala Leu 325 330 335 Gly Pro Gly Ala Thr Leu Glu Glu MetMet Val Ala Cys Gln Gly Val 340 345 350 Gly Gly Pro Thr His Lys Ala LysIle Leu Ala Glu Ala Met Ala Ser 355 360 365 Ala Gln Gln Asp Leu Lys GlyGly Tyr Thr Ala Val Phe Met Gln Arg 370 375 380 Gly Gln Asn Pro Asn ArgLys Gly Pro Ile Lys Cys Phe Asn Cys Gly 385 390 395 400 Lys Glu Gly HisIle Ala Lys Asn Cys Arg Ala Pro Arg Arg Arg Gly 405 410 415 Tyr Trp LysCys Gly Gln Glu Gly His Gln Met Lys Asp Cys Lys Asn 420 425 430 Gly ArgGln Ala Asn Phe Leu Gly Lys Tyr Trp Pro Pro Gly Gly Thr 435 440 445 ArgPro Ala Asn Tyr Val Gln Lys Gln Val Ser Pro Ser Ala Pro Pro 450 455 460Met Glu Glu Ala Val Lys Glu Gln Glu Asn Gln Asn Gln Lys Gly Asp 465 470475 480 Gln Glu Glu Leu Tyr Pro Phe Ala Ser Leu Lys Ser Leu Phe Gly Thr485 490 495 Asp Gln 61 35 PRT Human immunodeficiency virus type 1 61 ArgIle Leu Ala Val Glu Arg Tyr Leu Lys Asp Gln Gln Leu Leu Gly 1 5 10 15Ile Trp Gly Cys Ser Gly Lys Leu Ile Cys Thr Thr Ala Val Pro Trp 20 25 30Asn Ala Ser 35 62 35 PRT Human immunodeficiency virus 62 Arg Leu Gln AlaLeu Glu Thr Leu Ile Gln Asn Gln Gln Arg Leu Asn 1 5 10 15 Leu Trp GlyCys Lys Gly Lys Leu Ile Cys Tyr Thr Ser Val Lys Trp 20 25 30 Asn Thr Ser35 63 25 PRT Human immunodeficiency virus 63 Trp Gly Ile Arg Gln Leu ArgAla Arg Leu Gln Ala Leu Glu Thr Leu 1 5 10 15 Ile Gln Asn Gln Gln ArgLeu Asn Leu 20 25 64 22 DNA Artificial Sequence Description ofArtificial Sequence primer 64 ccataatatt cagcagaact ag 22 65 18 DNAArtificial Sequence Description of Artificial Sequence primer 65gctgattctg tataaggg 18 66 36 PRT Human immunodeficiency virus type 1 66Cys Thr Arg Pro Asn Asn Asn Thr Arg Lys Ser Ile Arg Ile Gln Arg 1 5 1015 Gly Pro Gly Arg Ala Phe Val Thr Ile Gly Lys Ile Gly Asn Met Arg 20 2530 Gln Ala His Cys 35 67 36 PRT Human immunodeficiency virus type 2 67Cys Lys Arg Pro Gly Asn Lys Ile Val Lys Gln Ile Met Leu Met Ser 1 5 1015 Gly His Val Phe His Ser His Tyr Gln Pro Ile Asn Lys Arg Pro Arg 20 2530 Gln Ala Trp Cys 35

1) an immunodeficiency virus of the HIV group, or variants of thisvirus, which exhibits the essential morphological and immunologicalproperties of the retrovirus which has the designation MVP-5180/91 andwhich has been deposited with the European collection of animal cellcultures (ECACC) under no: V 920 92
 318. 2) The immunodeficiency virusas claimed in claim 1, which exhibits a protein band in a Western blotwhich corresponds to reverse transcriptase and is 3-7 kilodaltonssmaller than the corresponding band of the HIV-1 and/or HIV-2 viruses.3) The immunodeficiency virus as claimed in one of claim 1 or 2, whichretrovirus exhibits less reactivity with a monoclonal antibody directedagainst protein p 24, related to reverse transcriptase activity, thandoes the HIV-1 virus, and more activity, related to the activity ofreverse transcriptase, than does HIV-2. 4) The immunodeficiency virus asclaimed in one of the preceding claims, wherein antigen/antibodyreactions with its transmembrane protein gp 41 are readily detectableusing sera from patients originating from Africa, and wherein only arelatively small antigen/antibody reaction, or no such reaction, can bedetected with the gp-41 using sera from patients originating fromGermany. 5) The immunodeficiency virus as claimed in one of theabovementioned claims, which has an RNA sequence which is homologous tothe extent of about 75% or more, based on the entire genome, with theRNA of the deposited virus. 6) The immunodeficiency virus as claimed inone of the abovementioned claims, which has an RNA sequence which ishomologous to the extent of at least 75% with the RNA sequence ofTable
 1. 7) The immunodeficiency virus as claimed in one of claims 1 to5, which has a nucleotide sequence which is homologous to the extent ofat least 75% with the sequence of Table 3, or parts thereof. 8) Theimmunodeficiency virus as claimed in claim 7, wherein the part of thesequence is at least 50 nucleotides long. 9) The immunodeficiency virusas claimed in one of the preceding claims, which has a sequence orconstituent sequence which corresponds to FIG. 4 or is homologous tothis sequence, where the differences from the sequence given in FIG. 4,related to the gene loci, are at most: LTR: 17%, GAG: 29%, POL: 25%,VIF: 31%, ENV: 46%, NEF: 16%. 10) The immunodeficiency virus as claimedin one of the preceding claims, which has a sequence or constituentsequence which corresponds to FIG. 4 or is homologous to this sequence,where the differences from the sequence given in FIG. 4, related to thegene loci, are at most: LTR: 10%, GAG: 14%, POL: 12%, VIF: 15%, ENV:22%, NEF: 10%. 11) cDNA which is complementary to the RNA, or partsthereof, of the immunodeficiency virus MVP-5180/91 deposited with theEuropean Collection of Animal Cell Cultures (ECACC) under No. V 920 92318, or of a virus as claimed in one of claims 1-10. 12) Recombinant DNAwhich contains cDNA as claimed in claim
 11. 13) An antigen which wasprepared using the cDNA as claimed in claim 11 or the recombinant DNA asclaimed in claim 12, or using the amino acid structure which can bededuced from its cDNA. 14) The antigen as claimed in claim 13, which isa protein or peptide. 15) The antigen as claimed in one of claim 13 or14, which has an amino acid sequence which corresponds to Table 3 or toa constituent sequence thereof. 16) The antigen as claimed in claim 15,wherein the constituent sequence has at least 10 amino acids. 17) Theantigen as claimed in claim 15, which has the amino acid sequenceRLQALETLIQNQQRLNLWGCKGKLICYTSVKWNTS, or a constituent sequence thereofhaving at least 6 consecutive amino acids. 18) An antigen which wasprepared from an immunodeficiency virus as claimed in one of claims 1 to10. 19) The antigen as claimed in one of claims 13 to 18, which wasprepared recombinantly. 20) The antigen as claimed in one of claims 13to 17, which was prepared synthetically. 21) A test kit for detectingantibodies against viruses which cause immuno deficiency, whereinantigen as claimed in claims 13 to 20 is employed. 22) The test kit asclaimed in claim 21, which is a Western blot. 23) The test kit asclaimed in claim 21, which is an ELISA test or a fluorescence-antibodydetection test. 24) Use of the immunodeficiency virus as claimed in oneof claims 1 to 10 and/or of the cDNA as claimed in claim 11 or 12 and/orof an antigen as claimed in claims 13 to 20 for detecting retroviruseswhich cause immune deficiency. 25) Use of a retrovirus as claimed in oneof claims 1 to 10, of a cDNA as claimed in claim 11 or 12 and/or of anantigen as claimed in claims 13 to 20 for preparing vaccines. 26)Ribonucleic acid characterized in that the ribonucleic acid is codingfor an immunodeficiency virus according to one of claims 1 to
 10. 27. Apeptide antigen comprising an amino acid sequence encoded by SEQ IDNO:56.
 28. The antigen of claim 27, wherein said amino acid sequence isat least 6 residues in length.
 29. The antigen of claim 27, wherein saidamino acid sequence is at least 10 residues in length.
 30. The antigenof claim 27, wherein said amino acid sequence is at least 15 residues inlength.
 31. The antigen of claim 27, wherein said amino acid sequence isat least 16 residues in length.
 32. The antigen of claim 27, whereinsaid amino acid sequence is at least 33 residues in length.
 33. Theantigen of claim 27, wherein said antigen comprises an amino acidsequence that is encoded by nucleotides 817-2310 of SEQ ID NO:56. 34.The antigen of claim 27, wherein said antigen comprises an amino acidsequence that is encoded by nucleotides 2073-5153 of SEQ ID NO:56. 35.The antigen of claim 27, wherein said antigen comprises an amino acidsequence that is encoded by nucleotides 6260-8887 of SEQ ID NO:56.
 36. Apeptide antigen comprising the amino acid sequence of SEQ ID NO:46. 37.A peptide antigen comprising an amino acid sequence present in SEQ IDNO:46.
 38. The antigen of claim 37, wherein said amino acid sequence isat least 6 residues in length.
 39. The antigen of claim 37, wherein saidamino acid sequence is at least 10 residues in length.
 40. The antigenof claim 37, wherein said amino acid sequence is at least 15 residues inlength.
 41. The antigen of claim 37, wherein said amino acid sequence isat least 16 residues in length.
 42. The antigen of claim 37, whereinsaid amino acid sequence is at least 33 residues in length.
 43. Apeptide antigen comprising the amino acid sequence of SEQ ID NO:39. 44.A peptide antigen comprising an amino acid sequence present in SEQ IDNO:39.
 45. The antigen of claim 44, wherein said amino acid sequence isat least 6 residues in length.
 46. The antigen of claim 44, wherein saidamino acid sequence is at least 10 residues in length.
 47. The antigenof claim 44, wherein said amino acid sequence is at least 15 residues inlength.
 48. The antigen of claim 44, wherein said amino acid sequence isat least 16 residues in length.
 49. The antigen of claim 44, whereinsaid amino acid sequence is at least 33 residues in length.
 50. Apeptide antigen comprising the amino acid sequence of SEQ ID NO:63. 51.A peptide antigen comprising an amino acid sequence present in SEQ IDNO:63.
 52. The antigen of claim 51, wherein said amino acid sequence isat least 6 residues in length.
 53. The antigen of claim 51, wherein saidamino acid sequence is at least 10 residues in length.
 54. The antigenof claim 51, wherein said amino acid sequence is at least 15 residues inlength.
 55. The antigen of claim 51, wherein said amino acid sequence isat least 16 residues in length.
 56. A peptide antigen comprising thesequence of SEQ ID NO:62.
 57. A peptide antigen comprising an amino acidsequence present in SEQ ID NO:62.
 58. The antigen of claim 57, whereinsaid amino acid sequence is at least 6 residues in length.
 59. Theantigen of claim 57, wherein said amino acid sequence is at least 10residues in length.
 60. The antigen of claim 57, wherein said amino acidsequence is at least 15 residues in length.
 61. The antigen of claim 57,wherein said amino acid sequence is at least 16 residues in length. 62.The antigen of claim 57, wherein said amino acid sequence is at least 33residues in length.
 63. A peptide antigen comprising the sequence of SEQID NO:59.
 64. A peptide antigen comprising an amino acid sequencepresent in SEQ ID NO:59.
 65. The antigen of claim 64, wherein said aminoacid sequence is at least 6 residues in length.
 66. The antigen of claim64, wherein said amino acid sequence is at least 10 residues in length.67. The antigen of claim 64, wherein said amino acid sequence is atleast 15 residues in length.
 68. The antigen of claim 64, wherein saidamino acid sequence is at least 16 residues in length.
 69. The antigenof claim 64, wherein said amino acid sequence is at least 33 residues inlength.
 70. A peptide antigen comprising the sequence of SEQ ID NO:60.71. A peptide antigen comprising an amino acid sequence present in SEQID NO:60.
 72. The antigen of claim 71, wherein said amino acid sequenceis at least 6 residues in length.
 73. The antigen of claim 71, whereinsaid amino acid sequence is at least 10 residues in length.
 74. Theantigen of claim 71, wherein said amino acid sequence is at least 15residues in length.
 75. The antigen of claim 71, wherein said amino acidsequence is at least 16 residues in length.
 76. The antigen of claim 71,wherein said amino acid sequence is at least 33 residues in length. 77.A peptide antigen comprising the amino acid sequence of SEQ ID NO:54.78. A peptide antigen comprising an amino acid sequence present in SEQID NO:54.
 79. The antigen of claim 78, wherein said amino acid sequenceis at least 6 residues in length.
 80. The antigen of claim 78, whereinsaid amino acid sequence is at least 10 residues in length.
 81. Theantigen of claim 78, wherein said amino acid sequence is at least 15residues in length.
 82. The antigen of claim 78, wherein said amino acidsequence is at least 16 residues in length.
 83. The antigen of claim 78,wherein said amino acid sequence is at least 33 residues in length. 84.A peptide antigen comprising the amino acid sequence of SEQ ID NO:55.85. A peptide antigen comprising an amino acid sequence present in SEQID NO:55.
 86. The antigen of claim 85, wherein said amino acid sequenceis at least 6 residues in length.
 87. The antigen of claim 85, whereinsaid amino acid sequence is at least 10 residues in length.
 88. Theantigen of claim 85, wherein said amino acid sequence is at least 15residues in length.
 89. The antigen of claim 85, wherein said amino acidsequence is at least 16 residues in length.
 90. The antigen of claim 85,wherein said amino acid sequence is at least 33 residues in length. 91.A peptide antigen comprising a peptide present in the GAG protein ofMvP-5180/91.
 92. A peptide antigen comprising a peptide present in thePOL protein of MvP-5180/91.
 93. A peptide antigen comprising a peptidepresent in the VIF protein of MvP-5180/91.
 94. A peptide antigencomprising a peptide present in the ENV protein of MvP-5180/91.
 95. Apeptide antigen comprising a peptide present in the NEF protein ofMvP-5180/91.